Relevant bibliographies by topics / In vitro culture

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  1. Journal articles
  2. Dissertations / Theses
  3. Books
  4. Book chapters
  5. Conference papers
  6. Reports

Academic literature on the topic 'In vitro culture'

Author: Grafiati
Published: 4 June 2021
Last updated: 6 July 2024

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Journal articles on the topic "In vitro culture"

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Nishii, R., K. Imai, H. Koyama, and O. Dochi. "166 EFFECT OF INDIVIDUAL CULTURE SYSTEM ON IN VITRO DEVELOPMENT OF IN VITRO-MATURED - IN VITRO-FERTILIZED BOVINE EMBRYOS." Reproduction, Fertility and Development 24, no. 1 (2012): 195. http://dx.doi.org/10.1071/rdv24n1ab166.

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An individual in vitro culture system for bovine embryo needs to be developed for the study of embryo developmental competence. The objective of this study was to examine the effect of individual culture systems on the development of in vitro-matured–in vitro-fertilized bovine embryos. Two individual culture systems were compared. Cumulus–oocyte complexes (COC) were collected by aspiration of ovarian follicles (diameter, 2 to 5 mm) obtained from a local abattoir. The COC were matured in TCM-199 supplemented with 5% calf serum and 0.02 AU mL–1 of FSH. Groups of 20 COC were incubated in 100-μL droplets of IVM media at 38.5°C under an atmosphere of 5% CO2 for 20 h. After 18 h of gamete co-culture (3 × 106 sperm mL–1), the presumptive zygotes were cultured in CR1aa medium supplemented with 5% calf serum at 38.5°C under an atmosphere of 5% CO2, 5% O2 and 90% N2 for 9 days (fertilization = Day 0). The presumptive zygotes were randomly assigned to 1 of the following 3 treatments: single culture (1 zygote was cultured in a 5-μL droplet), well-of-the-well (WOW; Sugimura et al. 2010 Biol. Reprod. 83, 970–978) culture (25 zygotes were cultured individually in each 125-μL droplet) and control culture (25 zygotes were cultured in a 125-μL droplet). Embryo development was evaluated for cleavage and blastocyst rates, on Day 2 and Day 7 to 9 after IVF, respectively. The rates of development up to the blastocyst stage and total cell number in the blastocysts, determined by an air-drying method, were investigated. The cleavage and blastocyst rates were analysed by the chi-square test and the total cell numbers were analysed by ANOVA. The cleavage rates were significantly higher in the control and WOW groups than in the single-culture group (P < 0.01) and the blastocyst rates were significantly lower in the single-culture group than in the control culture group (P < 0.05; Table 1). The total cell numbers (mean ± s.d.) of blastocysts did not significantly differ between the single culture (154.6 ± 21.8), control culture (155.2 ± 22.5) and WOW culture (159.8 ± 27.0) groups. These results indicate that although the blastocyst rate was lower in the single-culture system than in the WOW or group culture system, in vitro-matured–in vitro-fertilized bovine embryos can be cultured using the single-culture system. Moreover, the quality of blastocysts developed by the single-culture system is the same as that of blastocysts developed using the other 2 culture systems. Table 1.Effect of different culture methods for bovine embryo development
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TYNYKULOV, M. K., A. B. AKHMETOVA, A. B. ABZHALELOV, Sh E. ARYSTANOVA, A. U. UTAUBAYEVA, Zh N. UALIAKHMETOVA, А. U. TUYAKBAYEVA, and А. Zh ТEMIRKHANOV. "КУЛЬТУРА РАСТИТЕЛЬНЫХ КЛЕТОК IN VITRO AJUGA TURKESTANICA." МИКРОБИОЛОГИЯ ЖӘНЕ ВИРУСОЛОГИЯ, no. 2(45) (June 19, 2024): 159–80. http://dx.doi.org/10.53729/mv-as.2024.02.10.

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The article presents the growth rates of cells of new lines obtained by inoculation-strains of the culture of the endemic plant Ajuga turkestanica. Scientific research was conducted in 2022 in the laboratory of biology of cultured cells of the Institute of Plant Physiology named after K.A. Timiryazev. The purpose of the work: to obtain new tissue and cell culture lines in vitro of the endemic Ajuga turkestanica plant, as well as to study the possibility of regulating culture growth and biosynthesis of secondary metabolism products by the influence of various factors. The objects of the study are suspension cultures of Ajuga turkestanica cells. Source material: callus strain Ajuga turkestanica (Regel) Briq. At21, received in 2016, as well as ready-made frozen collection bases from the cryobank of the Timiryazev IGF of the Russian Academy of Sciences. Research methods: Obtaining a suspension of cells from callus tissue; cultivation of suspension cultures; determination of growth characteristics of the culture: the content of dry and raw biomass in a liter of medium, the concentration of cells in the medium, the viability of the culture. The results of the evaluation of the growth characteristics of suspension crops were obtained. Practical significance: the obtained suspension cultures can be used for various biotechnological studies, such as the production of biologically active substances, the study of secondary metabolism, the development of biotechnological methods for the production of medicines. В статье приведены ростовые показатели клеток новых полученных путем инокуляции линий-штаммов культуры эндемичного растения Ajuga turkestanica. Научные исследования проведены в 2022 году в лаборатории биологии культивируемых клеток Института физиологи растений им. К.А. Тимирязева. Цель работы: получение новых линий культуры тканей и клеток in vitro эндемичного растения Ajuga turkestanica, а также изучение возможности регуляции роста культуры и биосинтеза продуктов вторичного метаболизма воздействием различных факторов. Объектами исследованияявляются суспензионные культуры клеток Ajuga turkestanica. Исходный материал: каллусный штамм Ajuga turkestanica (Regel) Briq. Ат21, полученный в 2016 году, а также готовые замороженные коллекционные базы из криобанка ИФР им. К.А. Тимирязева РАН РФ. Методы исследования: получение суспензии клеток из каллусной ткани; культивирование суспензионных культур; определение ростовых характеристик культуры: содержание сухой и сырой биомассы в литре среды, концентрация клеток в среде, жизнеспособность культуры. Получены результаты по оценке ростовых характеристиках суспензионных культур. Практическая значимость: полученные суспензионные культуры могут быть использованы для различных биотехнологических исследований, таких как получение биологически активных веществ, изучение вторичного метаболизма, разработка биотехнологических методов производства лекарственных препаратов.
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Jarrahy, Reza, Weibiao Huang, George H. Rudkin, Jane M. Lee, Kenji Ishida, Micah D. Berry, Modar Sukkarieh, Benjamin M. Wu, Dean T. Yamaguchi, and Timothy A. Miller. "Osteogenic differentiation is inhibited and angiogenic expression is enhanced in MC3T3-E1 cells cultured on three-dimensional scaffolds." American Journal of Physiology-Cell Physiology 289, no. 2 (August 2005): C408—C414. http://dx.doi.org/10.1152/ajpcell.00196.2004.

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Osteogenic differentiation of osteoprogenitor cells in three-dimensional (3D) in vitro culture remains poorly understood. Using quantitative real-time RT-PCR techniques, we examined mRNA expression of alkaline phosphatase, osteocalcin, and vascular endothelial growth factor (VEGF) in murine preosteoblastic MC3T3-E1 cells cultured for 48 h and 14 days on conventional two-dimensional (2D) poly(l-lactide-co-glycolide) (PLGA) films and 3D PLGA scaffolds. Differences in VEGF secretion and function between 2D and 3D culture systems were examined using Western blots and an in vitro Matrigel-based angiogenesis assay. Expression of both alkaline phosphatase and osteocalcin in cells cultured on 3D scaffolds was significantly downregulated relative to 2D controls in 48 h and 14 day cultures. In contrast, elevated levels of VEGF expression in 3D culture were noted at every time point in short- and long-term culture. VEGF protein secretion in 3D cultures was triple the amount of secretion observed in 2D controls. Conditioned medium from 3D cultures induced an enhanced level of angiogenic activity, as evidenced by increases in branch points observed in in vitro angiogenesis assays. These results collectively indicate that MC3T3-E1 cells commit to osteogenic differentiation at a slower rate when cultured on 3D PLGA scaffolds and that VEGF is preferentially expressed by these cells when they are cultured in three dimensions.
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Colombo, Martina, Isa Mohammed Alkali, Sylwia Prochowska, and Gaia Cecilia Luvoni. "Fighting Like Cats and Dogs: Challenges in Domestic Carnivore Oocyte Development and Promises of Innovative Culture Systems." Animals 11, no. 7 (July 19, 2021): 2135. http://dx.doi.org/10.3390/ani11072135.

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In vitro embryo production in cats and dogs still presents some challenges, and it needs to be optimized to transfer efficient protocols to related wild, endangered species. While the chemical composition of culture media has been the focus of several studies, the importance of culture substrates for oocyte and embryo culture has often been neglected. Traditional in vitro systems, i.e., two-dimensional cultures, do not resemble the physiological environments where cells develop, and they may cause morphological and functional alterations to oocytes and embryos. More modern three-dimensional and microfluidic culture system better mimic the structure and the stimuli found in in vivo conditions, and they could better support the development of oocytes and embryos in vitro, as well as the maintenance of more physiological behaviors. This review describes the different culture systems tested for domestic carnivore reproductive cells along the years, and it summarizes their effects on cultured cells with the purpose of analyzing innovative options to improve in vitro embryo production outcomes.
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Thakur, Anirudh, G. S. Sidhu, and Harminder Singh. "In vitro multiplication of peach rootstocks." Indian Journal of Horticulture 80, no. 03 (September 25, 2023): 251–57. http://dx.doi.org/10.58993/ijh/2023.80.3.4.

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The leaf yellowing and senescence during micro-propagation reduces the shoot proliferation rates with every sub-culture in peach rootstock (Prunus spp.). The effect of culture media and supplements on the proliferation of cultures during the micro-propagation of two peach rootstocks ‘Sharbati’ and ‘Flordaguard’ was studied. In both Prunus genotypes, the proliferated cultures decreased from 77.3% and 67.0 % after the first sub-culture to 35.3 % and 27.3 % after the third sub-culture in ‘Sharbati’ and ‘Flordaguard’, respectively. QL medium significantly improved the proportion of proliferated cultures over MS, WPM, DKW and the modified MS media. The highest proliferated cultures (79.0% and 70.7%) and shoot number per culture (4.2 and 3.7) were recorded after the third subculture, in ‘Sharbati’ and ‘Flordaguard’, respectively with QL medium. Both the rootstocks varied in their response to iron chelates. The higher proliferation rates were obtained with MS medium by substituting Fe-EDTA (0.1 mM) with Fe-EDDHA (0.1-0.3 mM) in ‘Sharbati’ and with Fe-Na EDTA (0.1 mM) in ‘Flordaguard’. In‘Sharbati’, the highest proliferated cultures (44.3%) after the third subculture were observed with Fe-EDDHA (0.3 mM). In ‘Flordaguard’, the highest proportion of proliferated cultures (45%) after the third subculture was recorded with Fe-Na EDTA (0.1 mM). Adding silver nitrate (3 mM) improved the shoot proliferation rates to the highest levels (63% and 51%) after third subculture in ‘Sharbati’ and ‘Flordaguard’, respectively. The highest rooting (34.3% and 38.0%) and number of roots per shoot (2.2 and 2.5) were recorded with 3.75 mM of IBA in ‘Sharbati’ and ‘Flordaguard’, respectively.
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Gupta, P. S. P., H. S. Ramesh, B. M. Manjunatha, S. Nandi, and J. P. Ravindra. "Production of buffalo embryos using oocytes from in vitro grown preantral follicles." Zygote 16, no. 1 (February 2008): 57–63. http://dx.doi.org/10.1017/s096719940700442x.

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SummaryThe present study examines the use of buffalo preantral follicles as a source of oocytes for in vitro embryo production. Preantral follicles were isolated from abattoir-derived buffalo ovaries and were grown for 100 days in five different culture systems: (1) minimum essential medium (MEM); (2) coconut water; (3) MEM + ovarian mesenchymal cell (OMC) co-culture; (4) MEM + granulosa cell (GC) co-culture; or (5) MEM + cumulus cell (CC) co-culture. Low growth rates for the preantral follicles were observed when follicles were cultured in MEM or coconut water medium. Moderate growth rates were seen for OMC and GC co-cultures, and high rates of growth were observed when follicles were grown in CC co-culture. The survival of preantral follicles was low in the MEM culture (<25%), but was over 75% in the other culture systems. Oocytes were not recovered from the MEM group, while an oocyte recovery rate of 80–100% was observed when the follicles were cultured with coconut water/somatic cells. Transferable embryos could be produced only with the oocytes obtained from preantral follicles grown in the OMC and CC co-culture systems. This study demonstrates, for the first time, that it is possible to produce buffalo embryos by in vitro fertilization of oocytes derived from in vitro grown preantral follicles.
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Haselager, Marco, Eduard Perelaer, Arnon P. Kater, and Eric Eldering. "Development of a Novel Lymph Node-Based 3D Culture System Promoting Chronic Lymphocytic Leukemia Proliferation and Survival." Blood 136, Supplement 1 (November 5, 2020): 47–48. http://dx.doi.org/10.1182/blood-2020-141962.

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INTRODUCTION. Primary chronic lymphocytic leukemia (CLL) cells, despite originating from a proliferative disease, rapidly undergo apoptosis in vitro in absence of microenvironmental survival signals1. Although co-culture with stromal cells or the addition of soluble factors can increase and extend CLL survival, no system permits the long-term expansion of CLL cells in vitro2. The difficulties of mimicking a physiologic microenvironment supporting CLL cells hinder in vitro studies of proliferation, drug screens and prevent propagation of rare subclones. For other cancers, various types of 3D cultures have been introduced utilizing scaffolds, gels, spheroid cultures and fluidic systems, representing a more accurate representation of the in vivo microenvironment3. Unlike solid tumors, secondary lymphoid tissues where CLL cells proliferate in vivo, do not derive from a single stem cell progenitor. Developing an appropriate 3D in vitro culture system for CLL is of obvious importance and may contribute pathophysiological relevance to study long-term CLL proliferation and more accurate drug screening4,5. Within the field of CLL, attempts have focused on bone marrow stroma, but it may be biologically and clinically more relevant to investigate the lymph node niche as this is the critical site of CLL proliferation6. METHODS. Primary CLL cells were cultured in various 3D systems including hydrogels, hanging drop cultures and ultra-low attachment plates (ULA) plates in parallel to an optimal 2D system, consisting of the culture of primary CLL cells on a monolayer of CD40L-presenting fibroblasts (3T40) or 3T3 negative control fibroblasts. CLL cells were either cultured as PBMCs alone, with or without T cells, or co-cultured with 3T40 or primary lymph node fibroblasts. CLL cells were either stimulated directly with IL-2, IL-15, IL-21 and CpG and/or indirectly via a T cell stimulation of anti-CD3/CD28. RESULTS. After testing and comparing multiple systems for the in vitro culture of CLL cells, we optimized a novel CLL culture system utilizing ULA plates creating spheroids of PBMCs isolated from peripheral blood. Without the addition of soluble factors or stroma, primary CLL cells in the ULA 3D model could be maintained in culture for 6 weeks as opposed to 1 week in the 2D system. Aside from significantly promoting CLL survival, cultures could be expanded approximately 3-4-fold over a course of 6 weeks using the ULA 3D model. 3D cultures showed a more consistent and significantly increased CLL proliferation compared to 2D cultures, independent of IGHV mutation status, increasing the average proliferation index of 2.87 to 3.90 (n=10). Additionally, co-culture with LN-derived stromal cells further increased CLL proliferation, reaching a maximum of 8 generations (n=6) (Figure 1). Lastly, when PBMCs were stimulated with IL-2, IL-15, IL-21 and CpG, spheroids developed proliferation center-like structures after 4 weeks of culture. CONCLUSIONS. We established a lymph node-based 3D in vitro culture system for CLL leading to increased CLL proliferation and survival compared to 2D systems. The set-up allows long-term expansion of CLL cells in vitro, as well as formation of proliferation center-like structures. We are currently optimizing drug resistance studies, expansion of specific CLL subclones and performing competition experiments. References: 1. Hamilton et al., Mimicking the tumour microenvironment: three different co-culture systems induce a similar phenotype but distinct proliferative signals in primary chronic lymphocytic leukaemia cells, 2012. 2. Asslaber et al., Mimicking the microenvironment in chronic lymphocytic leukaemia - where does the journey go?, 2013. 3. Gurski et al., 3D Matrices for Anti-Cancer Drug Testing and Development, 2010. 4. Nunes et al., 3D tumor spheroids as in vitro models to mimic in vivo human solid tumors resistance to therapeutic drugs, 2019. 5. Aljitwai et al., A novel three-dimensional stromal-based model for in vitro chemotherapy sensitivity testing of leukemia cells, 2014. 6. Van Gent et al., In vivo dynamics of stable chronic lymphocytic leukemia inversely correlates with somatic hypermutation levels and suggest no major leukemic turnover in bone marrow, 2008. Disclosures Kater: Genentech: Research Funding; Abbvie: Research Funding; Roche: Research Funding; Janssen: Research Funding; Celgene: Research Funding. Eldering:Celgene: Research Funding; Janssen: Research Funding; Genentech: Research Funding.
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Adelberg, J., M. Kroggel, and J. Toler. "Physical Environment in vitro Affects Laboratory and Nursery Growth of Micropropagated Hostas." HortTechnology 10, no. 4 (January 2000): 754–57. http://dx.doi.org/10.21273/horttech.10.4.754.

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Hosta ×hybrid Tratt. `Blue Cadet' and Hosta tokudama Tratt. `Newberry Gold' were micropropagated in shaken liquid culture and on agar media, in conventional vessels and vessels modified for ventilation in vitro. Acclimatization under intermittent mist and growth in an outdoor nursery during the late spring and summer were monitored by dry weight analysis of sample plants every 4 days for a 60-day period (ex vitro growth). Results for `Newberry Gold' were 1) in vitro shoot growth was greater in liquid than agar culture, regardless of vessel; 2) shoots from agar or liquid culture grew at similar rates ex vitro; 3) ex vitro root growth was greater for liquid than agar cultured plants, regardless of vessel type. Results for `Blue Cadet' were 1) in vitro and ex vitro shoot growth was greater in liquid than agar culture regardless of vessel type and 2) ex vitro root growth was greatest for liquid cultured plants from conventional vessels. Ventilated vessels were generally beneficial for agar but not liquid culture. Benefits of liquid culture for micropropagation of Hosta found in vitro are at least maintained and sometimes enhanced during ex vitro growth in the mist bed and nursery.
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du Preez, Francois, Antoinette Paula Malan, and Pia Addison. "Potential of in vivo- and in vitro-cultured entomopathogenic nematodes to infect Lobesia vanillana (Lepidoptera: Tortricidae) under laboratory conditions." PLOS ONE 16, no. 8 (August 16, 2021): e0242645. http://dx.doi.org/10.1371/journal.pone.0242645.

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Entomopathogenic nematodes (EPNs) have been successfully applied as biological control agents against above ground and soil stages of insect pests. However, for commercial application, it is crucial to mass culture these nematodes using in vitro liquid culture technology, as it is not attainable when using susceptible insects as hosts. Lobesia vanillana (Lepidoptera: Tortricidae) is regarded a sporadic pest of wine grapes in South Africa. The in vivo- and in vitro-cultured South African EPNs, Steinernema yirgalemense and Steinernema jeffreyense (Rhabditida: Steinernematidae), were evaluated against larvae and pupae of L. vanillana in laboratory bioassays. For larvae, high mortality was observed for all treatments: In vitro-cultured S. yirgalemense (98%) performed better than S. jeffreyense (73%), while within in vivo cultures, there was no difference between nematode species (both 83%). No significant difference was detected between in vivo- and in vitro cultures of the same nematode species. The LD50 of the in vitro-cultured S. yirgalemense, was 7.33 nematodes per larva. Mortality by infection was established by dissecting L. vanillana cadavers and confirming the presence of nematodes, which was > 90% for all treatments. Within in vitro cultures, both S. yirgalemense and S. jeffreyense were able to produce a new cohort of infective juveniles from L. vanillana larvae. Pupae, however, were found to be considerably less susceptible to EPN infection. This is the first study on the use of EPNs to control L. vanillana. The relative success of in vitro-cultured EPN species in laboratory assays, without any loss in pathogenicity, is encouraging for further research and development of this technology.
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Dewir, Yaser Hassan, Abdulla Alsadon, Ahmed Ali Al-Aizari, and Mohaidib Al-Mohidib. "In Vitro Floral Emergence and Improved Formation of Saffron Daughter Corms." Horticulturae 8, no. 10 (October 20, 2022): 973. http://dx.doi.org/10.3390/horticulturae8100973.

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In vitro cormogenesis is a potential tool for improving saffron production under controlled conditions. In this study, the effects of explant type, culture type, and medium supplements on saffron daughter corm formation in vitro were assessed. Saffron flowers emerged 30 days after culture, and the sizes of in-vitro- and ex-vitro-produced flowers and stigmas were similar. In vitro daughter corm formation and the saffron life cycle was completed after 10 and 14 weeks of culture, respectively. Using in vitro intact corms was more effective for corm production than using apical buds. Compared with apical bud explants, mother corm explants produced more corms with a higher fresh weight and diameter. Compared with solid culture, liquid cultures using bioreactors provided corms with a higher fresh weight and diameter, regardless of explant type. An ebb and flow system provided the highest cormlet fresh weight and diameter but the fewest cormlets, whereas an immersion system provided more cormlets with a smaller size. Saffron apical buds cultured with salicylic acid at 75 mg L−1 or glutamine at 600 mg L−1 exhibited the highest cormlet diameter and fresh weight. These findings will improve the process of in vitro cormogenesis and the production of saffron under controlled conditions.
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Dissertations / Theses on the topic "In vitro culture"

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Mendes, Anabela Lindo dos Santos. "Cultura in vitro de embriões imaturos em ameixeira europeia." Master's thesis, Universidade de Évora, 1997. http://hdl.handle.net/10174/12550.

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Foram colhidos semanalmente, desde os 7 aos 70 dias após a polinização (DAP), frutos de Prunus domestica cv. ‘Rainha Cláudia Verde' provenientes de polinização livre. Com o objectivo de obter plântulas, foram extraídos os óvulos e/ou embriões e os eixos embrionários e testados três meios de cultura base, adicionando ou não diferentes tipos de reguladores de crescimento (auxina, giberelina e citocinina) e amino-ácidos (caseína hidrolisada 500 mg/l), tendo-se também variado a concentração de sacarose (2 e 10%). Foi ainda testada a técnica de perfuração da testa do óvulo e a influência do frio (30 dias a 4°C), na quebra de dormência dos embriões e eixos embrionários. Os resultados mostram que, aos 49 DAP quando comparámos a cultura do embrião dentro do óvulo perfurado, com a cultura do embrião isolado no meio de cuhura, verificámos que esta última conduziu a um melhor desenvolvimento do embrião e a percentagens de sobrevivência mais elevadas. Para os embriões no início da fase cotiledonar (49 DAP; PF1=11), o meio de cultura de Chée & Pool (1987) com 2% Sacarose +Caseína Hidrolisada (500 mg/1), demonstrou ser o mais adequado para o crescimento e sobrevivência dos embriões, quando comparado com os meios Nitsch & Nitsch (1969) e Stewart & Hsu (1977). Os embriões colocados neste meio de cultura apresentavam um comprimento médio inicial de 1,62 mm e atingiram em média os 5,31 mm, ao fim de 60 dias em cultura. No entanto, só foi possível obter plântulas a partir de embriões em plena fase cotiledonar (56 DAP; PF1=44). Neste estádio de desenvolvimento, o frio teve um efeito positivo tanto na percentagem de germinação como na qualidade das plântulas obtidas. Para os embriões em plena fase cotiledonar o meio de cultura Nitsch & Nitsch (1969) com 2% Sacarose +BAP (0,1mg/l.) +2,4-D (0,1mg/l), mostrou ser o mais adequado para a germinação, contudo, as plântulas obtidas apresentaram baixa qualidade. ### Abstract - The fruits of Prunus domestica cv. `Rainha Cláudia Verde' subject to opera pollination were picked weekly, from 7 to 70 days after pollination (DAP), with the aim of obtaining seedlings; the ovules and/or embryos and axis were extracted and placed in three different media with different concentrations of growth regulators (auxin, cytokinin and gibberellin), acid amino (casein hidrolysate 500 mg/l) and sacarose (2 and 10%). The ovule perforation technique and cold treatment (30 days at 4°C) were also applied to break down dormancy of the embryos and the embryonic axis. The results show that, compared to the ovule culture, the isolated embryo culture, at 49 DAP, gives a better embryo growth and a higher percentage of survival. The most adequate mediam for the growth and survival of the embryos at the early cotiledonar stage (49 DAP, PF1=11) proved to be the Chée & Pool medium (1987) with 2% sacarose+ Casein Hidrolysate (500 mg/l), as compared to the Nitsch & Nitsch medium (1969) and Stewart & Hsu medium (1977). The embryos that were placed in this medium initially had an average length of 1.62mm and after 60 days in the medium they had obtained an average length of 5.31mm. However, it was only possible to obtain seedlings from embryos at the cotiledonar stage (56 DAP, PF1=44). At this stage of development, the cold treatment had a positive effect, both for the percentage of germination and for the quality of the seedlings. For the embryos at the cotiledonar stage the Nitsch & Nitsch medium (1969) with 2% sacarose + BAP(0.1 mg/1)+ 2,4-D(0.1 mg/1) proved to be the most successful one for germination, however, the quality of these seedlings was poor.
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Langan, Laura. "Fish intestinal cultures for ecotoxicological studies : in vitro and primary culture models." Thesis, University of Plymouth, 2017. http://hdl.handle.net/10026.1/9486.

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Ecotoxicity testing of chemicals for environmental risk assessment is an area where a high number of vertebrates are used across a variety of industrial sectors. The application of the 3Rs in toxicity testing using fish address both the ethical and societal concerns around this issue in addition to the increasing legislative requests for the incorporation of animal alternatives. This thesis aims to highlight the potential of 3D cell culture models to "bridge the gap" between in vitro and in vivo screening procedures for testing of chemicals with the potential to persist or bioaccumulate thereby improving the predictive power of screening procedures. This thesis examines two alternative methods for their potential use as an intestinal based toxicokinetic tool for environmental risk assessment, utilising an in vitro fish cell line replacement tool (RTgutGC). In addition, for the first time a new intestinal primary cell culture based model was developed to address both intestine region specific response (pyloric, anterior, mid and posterior) and size related adaptability to toxins. Paramagnetic oximetry was used to measure oxygen content within 3D structures (spheroids) in order to better understand the microenvironment of these culture models. Using histology, immunohistochemistry, transepithelial electrical resistance (TEER), transmission electron microscopy (TEM), metabolic, fluorescence and gene expression assays, the comparability of this system to native intestinal response was established. Following exposure to carefully chosen environmental contaminants (Benzo[a]pyrene and Copper), the RTgutGC cell line demonstrated comparable responses to existing literature in terms of uptake, metabolism, DNA damage and the presence an equivalent saturable level. Primary enterocytes cultured on transwell inserts remained viable for upto six weeks, with permeability and metabolic activity comparable to native tissue (both in vitro and ex vivo). Taken in combination, these features of enterocytes represent a profile more closely representative of the intestine then the widely used "gut sac" method. With the potential advantages of incorporating complexity at differing levels (connective tissue layer, intestinal bacteria biome), the intestinal models described offer the potential to screen highly persistent toxins which may require prolonged incubation, in addition to the exploration of complex experimental designs which minimise animal usage (uptake, depuration, uptake). As a consequence, the models developed within this thesis significantly enrich the emerging fish based in vitro testing strategies.
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Czechowiak, Caty. "Culture in vitro de méristèmes de Pelargoniums." Lille 1, 1988. http://www.theses.fr/1988LIL10064.

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Contrairement à la production massive de boutures contribuant à augmenter l'extension des virus et bactéries (Xanthomonas), la culture de méristèmes de pélargonium ensemencés sur du milieu de base additionné de faibles quantités hormonales peuvent régénérer des plantes exemptes de maladies. Cependant en cas de trop fortes concentrations en hormones, le méristème peut produire un cal organogène générateur d'une dérive génétique. La méthode basée sur deux variétés: super rose (P. Hederaefolium) et topscore (P. Hortorum) a ensuite pu être généralisée.
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Nelson, T. J. "In vitro culture of Theobroma cacao L." Thesis, University of Nottingham, 1987. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.376391.

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Bishop, Rebecca Louise. "Pneumocystis carinii : approaches to in vitro culture." Thesis, London School of Hygiene and Tropical Medicine (University of London), 1996. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.244726.

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Latif, Sjafrul. "In vitro culture of ginger and macadamia." Thesis, Queensland University of Technology, 2000.

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Najm, Nour Addeen. "In vitro culture studies of tick cell lines." Diss., lmu, 2012. http://nbn-resolving.de/urn:nbn:de:bvb:19-146638.

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Tascan, Ayse. "In vitro liquid culture systems of Scutellaria species." Connect to this title online, 2007. http://etd.lib.clemson.edu/documents/1181251555/.

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Kaparakis, Georgios. "In vitro culture of pepper (Capsicum annuum L.)." Thesis, University of Nottingham, 1999. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.297989.

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Sivathanu, Vivek. "In vitro models for airway epithelial cell culture." Thesis, Massachusetts Institute of Technology, 2013. http://hdl.handle.net/1721.1/81726.

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Thesis (S.M.)--Massachusetts Institute of Technology, Dept. of Mechanical Engineering, 2013.
Cataloged from PDF version of thesis.
Includes bibliographical references (p. 40-41).
This work is about the development of a physiologically relevant model of the human airway. Various factors such as the cell model, physiochemical factors such as the cell substrate properties including its stiffness, shear stress, stretch, the air-liquid interface and the biochemical factors in the medium influence the biology of the cells. The aim of this work is to closely approximate conditions in an in vivo situation by engineering the above conditions in to the in vitro platform. An assay to introduce the cell substrate properties was developed in a glass bottomed petri dish type culture as well as a microfluidic device culture. The influence of the cell substrate on airway epithelial cell monolayer formation was investigated in detail by changing the stiffness of the substrate independently by changing the gel concentration, the gel formation pH and the height of the gel from a hard substrate. Further, we found that biochemical growth factors have a huge role in cell monolayer formation. A real-time measurement of monolayer integrity using electrical resistance measurements was developed. A shear stress application platform was developed and a stretch application platform was designed. The applications of such a platform with the inclusion of various physiologically relevant factors include the study of physiologic evolution of microbes such as the influenza virus.
by Vivek Sivathanu.
S.M.
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Books on the topic "In vitro culture"

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Bonga, J. M., and P. von Aderkas. In Vitro Culture of Trees. Dordrecht: Springer Netherlands, 1992. http://dx.doi.org/10.1007/978-94-015-8058-8.

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Declerck, Stéphane, J. André Fortin, and Désiré-Georges Strullu, eds. In Vitro Culture of Mycorrhizas. Berlin, Heidelberg: Springer Berlin Heidelberg, 2005. http://dx.doi.org/10.1007/b138925.

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Bonga, J. M. In vitro culture of trees. Dordrecht: Kluwer Academic Publishers, 1992.

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Stéphane, Declerck, Strullu D. G, and Fortin J. André, eds. In vitro culture of mycorrhizas. Berlin: Springer, 2005.

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R, Watson Ronald, ed. In vitro methods of toxicology. Boca Raton: CRC Press, 1992.

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Pierik, R. L. M. In vitro culture of higher plants. 3rd ed. Dordrecht: Kluwer Academic Publishers, 1997.

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M, Pierik R. L. In vitro culture of higher plants. Dordrecht: Kluwer Academic Publishers, 1997.

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M, Pierik R. L. In vitro culture of higher plants. 4th ed. Dordrecht: M. Nijhoff, 1997.

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M, Pierik R. L. In vitro culture of higher plants. Dordrecht: M. Nijhoff, 1987.

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Pierik, R. L. M. In Vitro Culture of Higher Plants. Dordrecht: Springer Netherlands, 1987. http://dx.doi.org/10.1007/978-94-009-3621-8.

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Book chapters on the topic "In vitro culture"

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Maggiulli, Roberta, Filippo Ubaldi, and Laura F. Rienzi. "Oocyte Insemination and Culture." In In Vitro Fertilization, 83–98. New York, NY: Springer New York, 2012. http://dx.doi.org/10.1007/978-1-4419-9848-4_6.

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Trounson, A. "Fertilization and Embryo Culture." In Clinical In Vitro Fertilization, 33–50. London: Springer London, 1989. http://dx.doi.org/10.1007/978-1-4471-1664-6_4.

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Gardner, David K., and Michelle Lane. "Sequential Media for Human Blastocyst Culture." In In Vitro Fertilization, 157–70. Cham: Springer International Publishing, 2019. http://dx.doi.org/10.1007/978-3-319-43011-9_16.

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Pierik, R. L. M. "Embryo culture." In In Vitro Culture of Higher Plants, 139–48. Dordrecht: Springer Netherlands, 1997. http://dx.doi.org/10.1007/978-94-011-5750-6_15.

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Pierik, R. L. M. "Embryo culture." In In Vitro Culture of Higher Plants, 139–48. Dordrecht: Springer Netherlands, 1997. http://dx.doi.org/10.1007/978-94-017-1854-7_15.

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Pierik, R. L. M. "Embryo culture." In In Vitro Culture of Higher Plants, 139–48. Dordrecht: Springer Netherlands, 1987. http://dx.doi.org/10.1007/978-94-009-3621-8_16.

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Gardner, David K., and Michelle Lane. "Embryo Culture Systems." In Handbook of In Vitro Fertilization, 205–44. Taylor & Francis Group, 6000 Broken Sound Parkway NW, Suite 300, Boca Raton, FL 33487-2742: CRC Press, 2017. http://dx.doi.org/10.1201/9781315157269-15.

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Mothersill, Carmel. "Human Thyroid Culture." In In Vitro Toxicity Testing Protocols, 25–31. Totowa, NJ: Humana Press, 1995. http://dx.doi.org/10.1385/0-89603-282-5:25.

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Mothersill, Carmel. "Human Esophageal Culture." In In Vitro Toxicity Testing Protocols, 75–79. Totowa, NJ: Humana Press, 1995. http://dx.doi.org/10.1385/0-89603-282-5:75.

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Yuan, Peng, Liying Yan, and Gary D. Smith. "Microfluidics for Gamete Manipulation and Embryo Culture." In In Vitro Fertilization, 213–25. Cham: Springer International Publishing, 2019. http://dx.doi.org/10.1007/978-3-319-43011-9_20.

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Conference papers on the topic "In vitro culture"

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Alatortseva, T. A. "Petchoa in vitro culture." In 2nd International Scientific Conference "Plants and Microbes: the Future of Biotechnology". PLAMIC2020 Organizing committee, 2020. http://dx.doi.org/10.28983/plamic2020.015.

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Yegorova, N. A., M. S. Zagorskaya, and O. V. Yakimova. "Culture medium for mint micropropagation in vitro." In CURRENT STATE, PROBLEMS AND PROSPECTS OF THE DEVELOPMENT OF AGRARIAN SCIENCE. Federal State Budget Scientific Institution “Research Institute of Agriculture of Crimea”, 2020. http://dx.doi.org/10.33952/2542-0720-2020-5-9-10-88.

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The influence of the culture medium composition on the development of explants at the second stage of clonal micropropagation of mint (Mentha canadensis L. K59(4n)) was studied in order to improve the in vitro propagation technique. It was shown that the maximum multiplication rate (11.5) was provided by MS medium supplemented with BAP (1.0 mg/L), IAA (0.5 mg/L) and 2% sucrose.
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Antsiferov, D. V., E. A. Lukjanova, and D. A. Ivasenko. "Blue honeysuckle introduction to in vitro culture." In 2nd International Scientific Conference "Plants and Microbes: the Future of Biotechnology". PLAMIC2020 Organizing committee, 2020. http://dx.doi.org/10.28983/plamic2020.025.

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Lee, Cynthia R., Mauro Alini, and James C. Iatridis. "Organ Culture System for Mechanobiology Studies of the Intervertebral Disc." In ASME 2004 International Mechanical Engineering Congress and Exposition. ASMEDC, 2004. http://dx.doi.org/10.1115/imece2004-61248.

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The development of in vitro models is critical for furthering understanding of the intervertebral disc and the development of disc regeneration/tissue engineering. An in vitro culture system targeted towards mechano-biology studies of the intervertebral disc (IVD) was built and validated using bovine coccygeal discs. Discs were maintained in culture for up to one week with and without vertebral endplates. Water content and glycosaminoglycan content were found to be stable and cells were metabolically active when cultured under a 5kg static load.
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Semenova, E. F., K. V. Vedernikova, and E. Yu Schetneva. "In vitro culture of Nonea pulla DC. seeds." In CURRENT STATE, PROBLEMS AND PROSPECTS OF THE DEVELOPMENT OF AGRARIAN SCIENCE. Federal State Budget Scientific Institution “Research Institute of Agriculture of Crimea”, 2020. http://dx.doi.org/10.33952/2542-0720-2020-5-9-10-95.

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Nonea pulla DC. is a promising perennial medicinal plant growing in the Crimea. Controlled in vitro cultivation of nonea seeds allows improving the up-to-date techniques of seedlings preparation. The conducted experiments confirmed the low germinating capacity of seeds (5–9 %). To increase this parameter and to speed up the introduction process, we investigated the Nonea pulla in vitro culture. The initial phases of germination were expectedly observed during seeds cultivation. The seed swelling, rupture of pericarp and seed hull, release of germ with cotyledons, dehiscence of cotyledons were detected. Moreover, in some cases, no subsequent development was observed. However, normal germs formed in 60% of cases. Seeds also sprouted without the prior cold stratification. For the following growth, plants required a relatively simple culture medium. The maximum development conditions were reached after 1.0–1.5 months of in vitro cultivation (26±2 °С, illuminance of 2000–3000 lux, 16-hour photoperiod).
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Uzelac, Branka, Dragana Stojičić, Snežana Budimir, Svetlana Tošić, Bojan Zlatković, Saša Blagojević, Branislav Manić, Mirjana Janjanin, and Violeta Slavkovska. "ESSENTIAL OILS AS POTENTIAL BIOCONTROL PRODUCTS AGAINST PLANT PATHOGENS AND WEEDS: IN VITRO CULTURE APPROACH." In XXVII savetovanje o biotehnologiji. University of Kragujevac, Faculty of Agronomy, 2022. http://dx.doi.org/10.46793/sbt27.345u.

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Secondary metabolism in plant plays a major role in the survival of the plant in its ecosystem, mediating the interaction of the plant with its environment. Plant bioactive compounds are biosynthesized as a defensive strategy of plants in response to natural perturbations. A number of biological effects have been associated with the main monoterpenoids detected in investigated Micromeria spp. and Clinopodium spp. essential oils. One alternative for the production of these prospective biocontrol products is in vitro plant tissue culture. Our data suggest that the metabolic potential of in vitro shoot cultures of selected species can be manipulated by varying in vitro culture conditions.
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Raizer, O. B., and O. N. Khapilina. "Culture in vitro of rare and endemic species of Allium (A. ledebourianum, A. altaicum)." In 2nd International Scientific Conference "Plants and Microbes: the Future of Biotechnology". PLAMIC2020 Organizing committee, 2020. http://dx.doi.org/10.28983/plamic2020.204.

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Rare and endemic Allium ledebourianum and Allium altaicum were introduced into the culture in vitro. When cultivated under conditions of slight osmotic stress, viable cultures of rare and endangered Allium species were obtained.
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Asadova, S. Sh. "Introduction of Stevia rebaudiana Bertoni in in vitro culture." In 2nd International Scientific Conference "Plants and Microbes: the Future of Biotechnology". PLAMIC2020 Organizing committee, 2020. http://dx.doi.org/10.28983/plamic2020.294.

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A cell culture obtained from explants of adult plants and aseptic seedlings of the Stevia rebaudiana Bertoni variety with different levels of ploidy, characterized by high speed, proliferation and ability to morphogenesis.
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Zhang, Chunqiu, Pengfei Wu, Xin Wang, and Lilan Gao. "Perfusion System for Cell-Scaffold Complex Culture in Vitro." In 2019 IEEE International Conference on Mechatronics and Automation (ICMA). IEEE, 2019. http://dx.doi.org/10.1109/icma.2019.8816526.

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Isakov, Igor, and Nadezhda Bokareva. "INTRODUCTION OF KARELIAN BIRCH TO THE CULTURE IN VITRO." In Modern problems of animal and plant ecology. FSBE Institution of Higher Education Voronezh State University of Forestry and Technologies named after G.F. Morozov, 2021. http://dx.doi.org/10.34220/mpeapw2021_5-9.

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At present, the biological diversity of tree species is drying up. One of the main reasons for extinction is the destructive anthropogenic impact. According to the latest data, it became known that the Karelian birch was included in the Red Book of the Republic of Karelia as an endangered and diminishing species. The in vitro clonal micropropagation technology can help to quickly restore the population of Karelian birch. And also the technology under consideration will help to massively produce seedlings and seedlings of Karelian birch for both decorative and silvicultural purposes.
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Reports on the topic "In vitro culture"

1

Labrune, Elsa, Bruno Salle, and Jacqueline Lornage. An update on in vitro folliculogenesis: a new technique for post-cancer fertility. INPLASY - International Platform of Registered Systematic Review and Meta-analysis Protocols, August 2022. http://dx.doi.org/10.37766/inplasy2022.8.0111.

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Review question / Objective: The present review intends to summarize the progress of in vitro folliculogenesis in humans. It focuses on the culture media and then, according to the culture stage, on the different culture systems developed with comments on the results obtained. Condition being studied: This review focuses on the progress of in vitro folliculogenesis in humans. Eligibility criteria: Inclusion criteria : all original English-language articles on in vitro folliculogenesis from ovarian tissue in humans; exclusion criteria: non-English papers, works on animals, in vitro maturation and in vivo maturation works carried out within the context of in vitro fertilization protocols, studies on in vitro folliculogenesis that checked slow freezing and/or vitrification of ovarian tissue, studies on frozen or vitrified tissues (these do not have the same objective), studies on short culture times, and studies that lacked major results.
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Grover, Paramjit, M. F. Rahman, and M. Mahboob. Bio-Physicochemical Interactions of Engineered Nanomaterials in In Vitro Cell Culture Model. Fort Belvoir, VA: Defense Technical Information Center, August 2012. http://dx.doi.org/10.21236/ada567065.

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Guzman, Juantia J., Clark L. Gross, William J. Smith, and Susan A. Kelly. In Vitro Cytotoxicity Assays of Human Epidermal Keratinocytes in Culture Exposed to Sulfur Mustard. Fort Belvoir, VA: Defense Technical Information Center, January 2000. http://dx.doi.org/10.21236/ada390628.

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Gray, Dennis, and Victor Gaba. Genotype, Explant and Growth Regulator Effects in the Determination of Adventitious Regeneratin in Curcurbits, in Aid of Genetic Transformation. United States Department of Agriculture, June 1992. http://dx.doi.org/10.32747/1992.7561060.bard.

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The objective of this study was to gain an understanding of the in vitro regeneration process in watermelon and melon to enable the development of genetic transformation systems. The objectives were met and additional progress, unplanned during the original proposal, was made. Organogenic regeneration in vitro was studied in both melon and watermelon. Genotype played a significant role in regeneration. In melon, epidermal cells were responsible for most regeneration. Methods to obtain in vitro-derived watermelon tetraploids, needed for seedless varieties, were developed. The culture systems were refined so that they could be routinely used for transformation. Particle guns were constructed and Agrobacterium strains were obtained to study the effect of transformation procedures on culture system performance, allowing refinement of transformation protocols. The culture systems were shown to enable the stable transformation of both crops, allowing their future use for insertion of agriculturally-important genes. In addition, we showed that shoot apical meristems might be suitable target tissue for transformation and allow a wider range of genotypes to be used, which is needed for crops as diverse as cucurbits.
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Semaan, Dima, and Linda Scobie. Feasibility study for in vitro analysis of infectious foodborne HEV. Food Standards Agency, September 2022. http://dx.doi.org/10.46756/sci.fsa.wfa626.

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Hepatitis E virus (HEV) is a member of the Hepeviridae family capable of infecting humans producing a range of symptoms from mild disease to kidney failure. Epidemiological evidence suggests that hepatitis E genotype III and IV cases may be associated with the consumption of undercooked pork meat, offal and processed products such as sausages [1]. A study carried out by the Animal Health and Veterinary Laboratories Agency (AHVLA), found hepatitis E virus contamination in the UK pork production chain and that 10% of a small sample of retail pork sausages were contaminated with the virus [2]. Furthermore, studies have confirmed the presence of HEV in the food chain and the foodborne transmission of Hepatitis E virus to humans [reviewed in 5]. Likewise, Scottish shellfish at retail [6] have also been found positive for HEV viral nucleic acid and some preliminary studies indicate that the virus is also detectable in soft fruits (L Scobie; unpublished data). There are current misunderstandings in what this data represents, and these studies have raised further questions concerning the infectivity of the virus, the processing of these foods by industry and the cooking and/or preparation by caterers and consumers. There are significant gaps in the knowledge around viral infectivity, in particular the nature of the preparation of food matrices to isolate the virus, and also with respect to a consistent and suitable assay for confirming infectivity [1,3]. Currently, there is no suitable test for infectivity, and, in addition, we have no knowledge if specific food items would be detrimental to cells when assessing the presence of infectious virus in vitro. The FSA finalised a comprehensive critical review on the approaches to assess the infectivity of the HEV virus which is published [3] recommending that a cell culture based method should be developed for use with food. In order to proceed with the development of an infectivity culture method, there is a requirement to assess if food matrices are detrimental to cell culture cell survival. Other issues that may have affected the ability to develop a consistent method are the length of time the virally contaminated sample is exposed to the cells and the concentration of the virus present. In most cases, the sample is only exposed to the cells for around 1 hour and it has been shown that if the concentration is less that 1x103 copies then infection is not established [3,5,10,11].
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Nasircilar, Ayse Gul. In vitro Clonal Propagation of Endemic Allium junceum Subs. tridentatum via Immature and Mature Embryo Culture Method. "Prof. Marin Drinov" Publishing House of Bulgarian Academy of Sciences, August 2021. http://dx.doi.org/10.7546/crabs.2021.08.18.

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Ulukapi, Kamile, and Ayse Gul Nasircilar. Mutation Breeding Protocol for Development of Drought-tolerant Genotypes in Phaseolus vulgaris L. Using in-vitro Embryo Culture Techniques. "Prof. Marin Drinov" Publishing House of Bulgarian Academy of Sciences, November 2020. http://dx.doi.org/10.7546/crabs.2020.11.10.

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Zlotkin, Eliahu, Shizuo G. Kamita, Nor Chejanovsky, and S. Maeda. Targeting of an Expressed Insect Selective Neurotoxin by its Recombinant Baculovirus: Pharmacokinetic and Pharmacodynamic Aspects. United States Department of Agriculture, July 1995. http://dx.doi.org/10.32747/1995.7571354.bard.

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Objectives: 1) Clarification of the mode of potentiation of an expressed insect selective neurotoxin (AaIT) by its recombinant baculovirus. 2) In vitro formation and/or modification of neuroactive polypeptides for the design of new improved recombinant baculoviruses. Results: 1) A combined utilization of bioassays, LM-cytochemistry, the highly resolutive EM immunogold and electrical recording from the CNS of baculovirus and AaIT - expressing – recombinant baculovirus infected larvae it has been shown that the recombinant virus potentiates the effect of the toxin. Potentiation is achieved through its continuous expression in the infected tracheal epithelia thus providing a: a) Local supply of freshly produced toxin in the vicinity of its traget sites; b) Translocation of the expressed toxin to the insect CNS. The latter exposes the recombinant toxin to new, critical, target sites which are inaccessible through the natural route of scorpion envenomation. 2) Subjecting a recombinant AaIT toxin to a newly designed system of random mutagenesis results in large numbers of new AaIT genes with amino acid substitutions. The new or modified toxin genes were inserted into a linear BmNPV expressed in silkworm cell culture and assayed on blowfly and silkworm larvae. Thus a system for mass formation and screening of neuroactive agents was developed. Contribution to agriculture: 1) Demonstration of the insecticidal mechanism, capacity and utility of the combination of neuroactive polypeptides and recombinant pathogens. 2) Development of a simple in vitro system for the formation and selection of new neuroactive polypeptides.
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Petitte, James, Hefzibah Eyal-Giladi, and Malka Ginsburg. The Study of Primordial Germ Cell Development as a Tool for Gene Transfer in Chickens. United States Department of Agriculture, October 1991. http://dx.doi.org/10.32747/1991.7561071.bard.

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The ability to introduce novel genetic material into the genome of commercial poultry has been impeded by a lack of kowledge regarding the origin in the early embryo of the target cell of interest, namely, the germ cell. Hence, this project investigated the emergence of primordial germ cells (PGCs) during the early development of the avian embryo to aid in efforts to produce transgenic poultry on a routine basis. The strategy was to introduce foreign DNA into the area of the unincubated embryo that is destined to give rise to the germ line. The objectives of this project were: 1) to identify and localize a subpopulation of cells in the early embryo which will give rise to PGCs, 2) to determine the best location and stage of development to transfer donor cells for efficient germline chimerism, and 3) to transfect donor cells to produce transgenic/germline chimeric embryos. We show that by using the monoclonal antibody SSEA-1 and by various cell culture techniques that germ cells appear to segregate from the somatic lineages at St. X., a process that is gradual and continues through St. XIV. Using microsurgical transplantation between quail and chick embryos, we demonstrated that the inner 1/3 of the area pellucida between states X-XII gives rise to about 2/3 of the germ cell population at the time of their residence in the germinal crescent. Because of the non-localized emergence of PGCs, attempts to introduce foreign DNA into clonal precursors of germ cells through liposome-mediated transfection yielded unacceptable levels of efficiency. However, through our investigation of germ cell origins, an in vitro model of germ cell differentiation was developed that could offer a means of determining the factors required for the long term culture of avian PGCs thereby providing a convenient means of manipulating the avian genome.
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Cohen, Jerry D., and Ephraim Epstein. Metabolism of Auxins during Fruit Development and Ripening. United States Department of Agriculture, August 1995. http://dx.doi.org/10.32747/1995.7573064.bard.

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Abstract:
We had proposed to look at several aspects of auxin metabolism in fruit tissues: 1) IAA biosynthesis from tryptophan and IAA biosynthesis via the non-tryptophan pathway; 2) changes in the capacity to form conjugates and catabolites of auxin at different times during fruit development and; 3) the effects of modifying auxin metabolism in fruit tissues. The latter work focused primarily on the maize iaglu gene, with initial studies also using a bacterial gene for hydrolysis of IAA-aspartate. These metabolic and molecular studies were necessary to define potential benefits of auxin metabolism modification and will direct future efforts for crop improvement by genetic methods. An in vitro system was developed for the production of tomato fruit in culture starting from immature flowers in order to ascertain the effect of auxin modification on fruit ripening. IAA supplied to the fruit culture media prior to breaker stage resulted in an increase in the time period between breaker and red-ripe stages from 7 days without additional IAA to 12 days when 10-5 M IAA was added. These results suggest that significant changes in the ripening period could be obtained by alteration of auxin relationships in tomato fruit. We generated transgenic tomato plants that express either the maize iaglu gene or reduced levels of the gene that encodes the enzyme IAA-glucose synthetase. A modified shuttle vector pBI 121 expressing the maize iaglu gene in both sense and antisense orientations under a 35S promoter was used for the study. The sense plants showed total lack of root initiation and development. The antisense transgenic plants, on the other hand, had unusually well developed root systems at early stages in development. Analysis showed that the amount and activity of the endogenous 75 kDa IAGLU protein was reduced in these plants and consequently these plants had reduced levels of IAA-glucose and lower overall esterified IAA.
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