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Zeitschriftenartikel zum Thema "Cell culture techniques – laboratory manuals"

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Clothier, Richard H. „Book Review: Culture of Animals Cells: Manual of Basic Technique — Third Edition“. Alternatives to Laboratory Animals 23, Nr. 1 (Januar 1995): 161. http://dx.doi.org/10.1177/026119299502300121.

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Hulme, Lesley. „Book Review: Culture of Animal Cells: A Manual of Basic Technique— 2Nd Edition“. Alternatives to Laboratory Animals 16, Nr. 1 (September 1988): 97. http://dx.doi.org/10.1177/026119298801600119.

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Braylan, Raul C., und Elaine Kay Jordan. „Rapid and Simple Procedure For Isolation and Concentration Of Human Megakaryocytes From Marrow Aspirates“. Blood 122, Nr. 21 (15.11.2013): 5269. http://dx.doi.org/10.1182/blood.v122.21.5269.5269.

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Abstract Background Numerical and morphologic abnormalities of megakaryocytes (MKs) are present in a variety of primary or secondary bone marrow (BM) disorders such as myeloproliferative neoplasms, myelodysplastic syndromes or ITP. Currently, these changes are assessed exclusively on microscopic preparations of marrow aspirates and biopsies and are used as criteria for disease diagnosis, classification and therapy monitoring despite the inherent subjectivity of microscopic evaluations. In contrast to other cells, adequate studies of freshly isolated MKs have proven difficult because of the relative rarity of these cells in the BM (∼0.05% of total nucleated cells) and separation techniques such as density gradients, magnetic beads, centrifugal elutriation or fluorescence activated cell sorting are labor-intensive, time-consuming or costly. The primary means for studying MKs is based on the isolation of progenitors primed with cytokines to differentiate in culture. While extremely valuable, these techniques are not directly translatable to the routine clinical arena for assessing MK pathology in human BM. Mature marrow MKs are large, polyploid cells whose size distribution overlaps minimally with that of all other marrow cells. This distinct size threshold is a discriminatory parameter for MK isolation. Thus, we developed a simple and inexpensive manual mesh filtration method for separation of MKs that allows a rapid and easy, size-based concentration and purification of these cells. Methods We examined 15 discarded anonymized BM aspirate samples suitable for our analysis. These samples were from patients with a variety of hematologic disorders originally submitted to our laboratory for evaluation. Sample age ranged from 1-4 days (mean 1.5 days). Samples were diluted with heparin/albumin-containing buffered saline solution. The cell suspensions were first filtered through a 70µm filter to remove large BM stromal particles and then twice (by gravity) through 9mm-diameter wetted 8 µm nylon filters placed in a simple plastic holder. This filtration procedure allowed the retention of MKs and removal of smaller cells. The MK-rich suspension was collected by rinsing and flushing the filters with the buffered solution. On average, the procedure only lasted 15 minutes. The pre-filtered, 8 µm filter-retained, and effluent cell suspensions collected were stained with an FITC-anti CD61 or PE-anti CD41a antibody [Becton Dickinson (BD)] to label MKs, concurrently with the cell permeable DNA-binding DRAQ5 (Cell Signaling Technology) to identify all nucleated cells. Final cell viability was assessed by C12Resazurin (Molecular probes). Cell analysis was performed by flow cytometry (FCM) using a CANTO II flow cytometer (BD). Absolute MK enumeration was performed using the Flow Cytometry Absolute Count Standard beads (Bangs). MK enrichment efficiency was expressed as the percentage of MKs of all nucleated cells determined by FCM. No cell lysing, fixation or centrifugation was used at any step of the MK separation procedure. Results The median (range) volume of the BM aspirate samples used in this study was 0.54 (0.24-1.99) mL. MKs were identified by FCM on the basis of their large size, expression of platelet-associated antigens and DNA ploidy levels. The median (range) MK recovery was 31 (14-100) % of the original number of MKs and the yield was 9,882 (1,519-49,921) MKs per mL of BM aspirate. The median (range) fraction of MKs among all nucleated cells after filtration was 39 (14-68) %, representing a 904 (439-3029)-fold MK enrichment. The MK viability after filtration was near 100%. Conclusions This simple, gentle, rapid and inexpensive isolation and concentration method results in a MK recovery and purification that is comparable or better than other more elaborate techniques. Despite the inherent heterogeneity of the samples used, we obtained a reasonably good recovery of MKs per mL of marrow aspirate and more than 900-fold median MK concentration. The yield and level of purification of freshly isolated MKs obtained by this simple procedure may be useful in studies of a variety of primary or secondary marrow disorders. In particular, it should facilitate the application of analytical methods such as flow cytometry or in situ hybridization, and even be useful for biochemical or molecular testing that require adequate cell representation and purity. Disclosures: No relevant conflicts of interest to declare.
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Hidayat, Nurul, Mega Novia Putri und Ronal Kurniawan. „Nannochloropsis sp phytoplankton culture technique laboratory scale“. South East Asian Marine Sciences Journal 1, Nr. 2 (15.03.2024): 73–76. http://dx.doi.org/10.61761/seamas.1.2.73-76.

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Nannochloropsis sp is a phytoplankton often used in marine fish hatchery activities as feed for the mass production of rotifers, and its availability is very much needed for rearing marine fish larvae. This activity aims to study pure culture techniques for Nannochloropsis sp on a laboratory scale. The method used is a literature study method and a direct practical method regarding Nannochloropsis sp phytoplankton culture techniques laboratory scale and conducted interviews with employees at the BPBL Batam Phytoplankton Live Food Production Unit Laboratory. Based on observations, it was found that the peak growth or optimum cell density of Nannochloropsis sp. occurred on the sixth day in a 1000 mL Erlenmeyer, namely 60.95 - 62.10 million cells/mL with an initial density of 10.15 - 10.55 million cells/mL and in a 2000 mL Erlenmeyer, namely 58.75 - 60, 25 million cells/mL with an initial density of 8.25 – 8.45 million cells/mL.
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Lewis, Jennifer R., Mark S. Kotur, Omar Butt, Sumant Kulcarni, Alyssa A. Riley, Nick Ferrell, Kathryn D. Sullivan und Mauro Ferrari. „Biotechnology Apprenticeship for Secondary-Level Students: Teaching Advanced Cell Culture Techniques for Research“. Cell Biology Education 1, Nr. 1 (März 2002): 26–42. http://dx.doi.org/10.1187/cbe.02-02-0003.

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The purpose of this article is to discuss small-group apprenticeships (SGAs) as a method to instruct cell culture techniques to high school participants. The study aimed to teach cell culture practices and to introduce advanced imaging techniques to solve various biomedical engineering problems. Participants designed and completed experiments using both flow cytometry and laser scanning cytometry during the 1-month summer apprenticeship. In addition to effectively and efficiently teaching cell biology laboratory techniques, this course design provided an opportunity for research training, career exploration, and mentoring. Students participated in active research projects, working with a skilled interdisciplinary team of researchers in a large research institution with access to state-of-the-art instrumentation. The instructors, composed of graduate students, laboratory managers, and principal investigators, worked well together to present a real and worthwhile research experience. The students enjoyed learning cell culture techniques while contributing to active research projects. The institution's researchers were equally enthusiastic to instruct and serve as mentors. In this article, we clarify and illuminate the value of small-group laboratory apprenticeships to the institution and the students by presenting the results and experiences of seven middle and high school participants and their instructors.
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Bleotu, Coralia, Carmen Diaconu, Mihaela Chivu, Irina Alexiu, Simona Ruta und Costin Cernescu. „Evaluation of TV cell line viral susceptibility using conventional cell culture techniques“. Open Medicine 1, Nr. 1 (01.03.2006): 12–22. http://dx.doi.org/10.2478/s11536-006-0007-x.

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AbstractDespite the fact that a lot of methods have been developed for rapid virus detection, classic cell culture is still “the golden standard”. The range of viruses that can be isolated and cultured in cell line systems is often limited by the susceptibility of cells to support viral replication. Since the primary cell culture, the best cellular system available to support replication of a large number of viruses, is very expensive and diffcult to obtain, cell lines, which are easier to manipulate, are commonly used for virus growth and isolation.In two previous papers we described the TV cell line initiated by our team from a laryngeal tumor, which harbors human papillomavirus (HPV) gene sequences. In this paper we analyze its capacity to support virus replication. Depending on the virus, different cytopathic effects were produced. Comparison of viral effect observed on this cell line with the effect obtained on other cell lines has been performed. This cell line might be used in the clinical virology laboratory for virus isolation.
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Ciapetti, Gabriela, Elisabetta Cenni, Daniela Cavedagna, Loredana Pratelli und Arturo Pizzoferrato. „Cell Culture Methods to Evaluate the Biocompatibility of Implant Materials“. Alternatives to Laboratory Animals 20, Nr. 1 (Januar 1992): 52–60. http://dx.doi.org/10.1177/026119299202000107.

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Cell culture techniques are usually used in the field of biomaterials research and development in order to detect toxic components. Morphological assays are the most widely used methods and give the very first information about the biological compatibility of materials. Cell function assays give more quantitative data, but the comparison of data between different laboratories is difficult. Some of the cell culture methods that are used for biocompatibility studies are described briefly here, and results from our laboratory are reported. Despite some inherent limitations of the cell culture techniques, they are an accurate and reliable method of predicting the biological compatibility of materials to be implanted in vivo.
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Marion, Rebecca E., Grant E. Gardner und Lisa D. Parks. „Multiweek cell culture project for use in upper-level biology laboratories“. Advances in Physiology Education 36, Nr. 2 (Juni 2012): 154–57. http://dx.doi.org/10.1152/advan.00080.2011.

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This article describes a laboratory protocol for a multiweek project piloted in a new upper-level biology laboratory (BIO 426) using cell culture techniques. Human embryonic kidney-293 cells were used, and several culture media and supplements were identified for students to design their own experiments. Treatments included amino acids, EGF, caffeine, epinephrine, heavy metals, and FBS. Students researched primary literature to determine their experimental variables, made their own solutions, and treated their cells over a period of 2 wk. Before this, a sterile technique laboratory was developed to teach students how to work with the cells and minimize contamination. Students designed their experiments, mixed their solutions, seeded their cells, and treated them with their control and experimental media. Students had the choice of manipulating a number of variables, including incubation times, exposure to treatment media, and temperature. At the end of the experiment, students observed the effects of their treatment, harvested and dyed their cells, counted relative cell numbers in control and treatment flasks, and determined the ratio of living to dead cells using a hemocytometer. At the conclusion of the experiment, students presented their findings in a poster presentation. This laboratory can be expanded or adapted to include additional cell lines and treatments. The ability to design and implement their own experiments has been shown to increase student engagement in the biology-related laboratory activities as well as develop the critical thinking skills needed for independent research.
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Shyam, Rohin, L. Reddy und Arunkumar Palaniappan. „Fabrication and Characterization Techniques of In Vitro 3D Tissue Models“. International Journal of Molecular Sciences 24, Nr. 3 (18.01.2023): 1912. http://dx.doi.org/10.3390/ijms24031912.

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The culturing of cells in the laboratory under controlled conditions has always been crucial for the advancement of scientific research. Cell-based assays have played an important role in providing simple, fast, accurate, and cost-effective methods in drug discovery, disease modeling, and tissue engineering while mitigating reliance on cost-intensive and ethically challenging animal studies. The techniques involved in culturing cells are critical as results are based on cellular response to drugs, cellular cues, external stimuli, and human physiology. In order to establish in vitro cultures, cells are either isolated from normal or diseased tissue and allowed to grow in two or three dimensions. Two-dimensional (2D) cell culture methods involve the proliferation of cells on flat rigid surfaces resulting in a monolayer culture, while in three-dimensional (3D) cell cultures, the additional dimension provides a more accurate representation of the tissue milieu. In this review, we discuss the various methods involved in the development of 3D cell culture systems emphasizing the differences between 2D and 3D systems and methods involved in the recapitulation of the organ-specific 3D microenvironment. In addition, we discuss the latest developments in 3D tissue model fabrication techniques, microfluidics-based organ-on-a-chip, and imaging as a characterization technique for 3D tissue models.
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Malovrh, Tadej, und Peter Hostnik. „Diagnostics procedures in rabies“. Veterinarski glasnik 59, Nr. 1-2 (2005): 99–105. http://dx.doi.org/10.2298/vetgl0502099m.

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Rabies is a major zoonosis for which diagnostic techniques can only be performed in the laboratory. Laboratory techniques are preferably oriented on tissue removed from the cranium: hippocampus (Ammon's horn), cerebellum and the medulla oblongata or tissue liquids. Clinical observation may only lead to a suspicion of rabies. The only way to perform a reliable diagnosis of the disease is to identify the virus or some of its specific components using laboratory tests such as histological identification of characteristic cell lesions, immunochemical identification of rabies virus antigen and virus isolation. Serological tests are rarely used in epidemiological surveys but much more frequently in control of the vaccination programs (e.g. oral vaccination). Most commonly used serological tests are the virus neutralization test on cell culture (FAVN), virus neutralization in mice and ELISA.
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Bücher zum Thema "Cell culture techniques – laboratory manuals"

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M, Clynes, Hrsg. Animal cell culture techniques. Berlin: Springer, 1998.

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1939-, Rae Ian F., Hrsg. General techniques of cell culture. Cambridge: Cambridge University Press, 1997.

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Mitry, Ragai R., und Robin D. Hughes. Human cell culture protocols. 3. Aufl. New York: Humana Press, 2012.

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Joanna, Picot, Hrsg. Human cell culture protocols. 2. Aufl. Totowa, N.J: Humana Press, 2005.

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D, Helgason Cheryl, und Miller Cindy L, Hrsg. Basic cell culture protocols. 3. Aufl. Totowa, N.J: Humana Press, 2005.

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Loyola-Vargas, Victor M., und Neftalí Ochoa-Alejo. Plant cell culture protocols. 3. Aufl. New York: Springer, 2012.

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Cree, Ian A. Cancer cell culture: Methods and protocols. 2. Aufl. Totowa, N.J: Humana Press, 2011.

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Haycock, John W. 3D cell culture: Methods and protocols. New York: Humana Press/Springer, 2011.

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Haycock, John W. 3D cell culture: Methods and protocols. New York: Humana Press/Springer, 2011.

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Randell, Scott H., und M. Leslie Fulcher. Epithelial cell culture protocols. 2. Aufl. New York: Humana Press, 2012.

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Buchteile zum Thema "Cell culture techniques – laboratory manuals"

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Jain, Anvi, Aaru Gulati, Khushi R. Mittal und Shalini Mani. „Cell Culture Laboratory“. In Techniques in Life Science and Biomedicine for the Non-Expert, 11–52. Cham: Springer International Publishing, 2023. http://dx.doi.org/10.1007/978-3-031-19485-6_2.

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Ganesh, Goutham V., Kannan Harithpriya, Dhamodharan Umapathy, Md Enamul Hoque und K. M. Ramkumar. „Laboratory Safety“. In Advanced Mammalian Cell Culture Techniques, 11–14. Boca Raton: CRC Press, 2023. http://dx.doi.org/10.1201/9781003397755-4.

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Seidman, Lisa A. „Common Calculations Relating to Animal Cell Culture Techniques“. In Basic Laboratory Calculations for Biotechnology, 391–412. 2. Aufl. Boca Raton: CRC Press, 2021. http://dx.doi.org/10.1201/9780429282744-26.

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Mani, Shalini. „Managing Sterility in Animal Cell Culture Laboratory“. In Techniques in Life Science and Biomedicine for the Non-Expert, 65–75. Cham: Springer International Publishing, 2023. http://dx.doi.org/10.1007/978-3-031-19485-6_4.

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Yukta, Kumari, Mansi Agarwal, Mekhla Pandey, Khushi Mittal, Vidushi Srivastava und Shalini Mani. „Good Laboratory Practices in Animal Cell Culture Laboratory and Biosafety Measures“. In Techniques in Life Science and Biomedicine for the Non-Expert, 53–64. Cham: Springer International Publishing, 2023. http://dx.doi.org/10.1007/978-3-031-19485-6_3.

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„Introduction to the Cell Culture Laboratory“. In Manuals in Biomedical Research, 11–22. WORLD SCIENTIFIC, 2010. http://dx.doi.org/10.1142/9789812834782_0002.

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„Introduction to the Cell Culture Laboratory“. In Manuals in Biomedical Research, 11–22. WORLD SCIENTIFIC, 2004. http://dx.doi.org/10.1142/9789812791122_0002.

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„Reagents and Techniques for Cell Culture“. In Manuals in Biomedical Research, 97–126. WORLD SCIENTIFIC, 2010. http://dx.doi.org/10.1142/9789812834782_0004.

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„Reagents and Techniques for Cell Culture“. In Manuals in Biomedical Research, 75–136. WORLD SCIENTIFIC, 2004. http://dx.doi.org/10.1142/9789812791122_0004.

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Jiabei, Tong, Thilakavathy Karuppiah, Sun Zhong, Akon Higuchi und Suresh Kumar Subbiah. „Protocols in stem cell culture“. In Stem Cell Laboratory Techniques, 41–69. Elsevier, 2023. http://dx.doi.org/10.1016/b978-0-12-823729-8.00006-3.

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Konferenzberichte zum Thema "Cell culture techniques – laboratory manuals"

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Zavrel, Erik A., Michael L. Shuler und Xiling Shen. „A Simple Aspect Ratio Dependent Method of Patterning Microwells for Selective Cell Attachment“. In 2018 Design of Medical Devices Conference. American Society of Mechanical Engineers, 2018. http://dx.doi.org/10.1115/dmd2018-6811.

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3-D culture has been shown to provide cells with a more physiologically authentic environment than traditional 2-D (planar) culture [1, 2]. 3-D cues allow cells to exhibit more realistic functions and behaviors, e.g., adhesion, spreading, migration, metabolic activity, and differentiation. Knowledge of changes in cell morphology, mechanics, and mobility in response to geometrical cues and topological stimuli is important for understanding normal and pathological cell development [3]. Microfabrication provides unique in vitro approaches to recapitulating in vivo conditions due to the ability to precisely control the cellular microenvironment [4, 5]. Microwell arrays have emerged as robust alternatives to traditional 2D cell culture substrates as they are relatively simple and compatible with existing laboratory techniques and instrumentation [6, 7]. In particular, microwells have been adopted as a biomimetic approach to modeling the unique micro-architecture of the epithelial lining of the gastrointestinal (GI) tract [8–10]. The inner (lumen-facing) surface of the intestine has a convoluted topography consisting of finger-like projections (villi) with deep well-like invaginations (crypts) between them. The dimensions of villi and crypts are on the order of hundreds of microns (100–700 μm in height and 50–250 μm in diameter) [11]. While microwells have proven important in the development of physiologically realistic in vitro models of human intestine, existing methods of ensuring their surface is suitable for cell culture are lacking. Sometimes it is desirable to selectively seed cells within microwells and confine or restrict them to the microwells in which they are seeded. Existing methods of patterning microwells for cell attachment either lack selectivity, meaning cells can adhere and migrate anywhere on the microwell array, i.e., inside microwells or outside of them, or necessitate sophisticated techniques such as micro-contact printing, which requires precise alignment and control to selectively pattern the bottoms of microwells for cell attachment [12, 13].
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Temenoff, Johnna S. „A Modular System to Examine Fibroblastic Differentiation of Mesenchymal Stem Cells Under Tensile Loading in Response to Changes in the Extracellular Environment“. In ASME 2011 Summer Bioengineering Conference. American Society of Mechanical Engineers, 2011. http://dx.doi.org/10.1115/sbc2011-53704.

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Hundreds of thousands of injuries to ligaments, tendons or the joint capsule occur in the U.S. each year, resulting in significant reduction of quality of life for many patients [1]. Existing reconstruction techniques for torn tendons/ligaments result in significant morbidity and cannot fully recapitulate the native joint biomechanics, leading to secondary degeneration over time, such as premature osteoarthritis. Thus, tissue-engineered alternatives to current grafts, potentially using stem cells in combination with an appropriate scaffold, are greatly needed. In response, our laboratory is investigating a novel hydrogel system and a custom tensile bioreactor as an in-vitro model to study the formation of both fibrous (ligament) tissue and the ligament-bone interface. In these studies, we examine the effect of tensile loading and the degradability of the surrounding environment on cellular morphology and tendon/ligament extracellular matrix (ECM) production by mesenchymal stem cells (MSCs). In particular, the response of MSCs embedded within hydrogels with varying degrees of susceptibility to degradation by collagenase is explored. In addition, proof-of-principle experiments are presented to extend this system to examine the effect of co-culture of multiple cell types on differentiation of MSCs in a milieu that mimics the bone-ligament insertion.
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Berichte der Organisationen zum Thema "Cell culture techniques – laboratory manuals"

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Corscadden, Louise, und Anjali Singh. Methods Of Cleaning And Sterilization. Maze Engineers, Dezember 2022. http://dx.doi.org/10.55157/cs20221207.

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Microbiology, tissue culture, medical, equipment manufacturing labs, and many research labs and industries need strict sterile environments for their diverse operations. Experiments, specifically those involving cell lines or microorganisms need to be conducted in a controlled environment. Contamination not only voids experiments, but also wastes effort, time, and money and when involving patients, it poses serious health risks. It is essential to be well-versed in laboratory sterilization techniques.
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