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Journal articles on the topic 'Organotypic'

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Author: Grafiati

Published: 1 April 2023

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1

Oh, Ji Won, Tsai-Ching Hsi, Christian Fernando Guerrero-Juarez, Raul Ramos, and Maksim V. Plikus. "Organotypic Skin Culture." Journal of Investigative Dermatology 133, no. 11 (November 2013): 1–4. http://dx.doi.org/10.1038/jid.2013.387.

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2

Froeling, Fieke E. M., John F. Marshall, and Hemant M. Kocher. "Pancreatic cancer organotypic cultures." Journal of Biotechnology 148, no. 1 (July 1, 2010): 16–23. http://dx.doi.org/10.1016/j.jbiotec.2010.01.008.

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3

Zieske, James, Eui‐Hong Chung, Xiaoqing Guo, and Audrey Hutcheon. "Human Corneal Organotypic Cultures." Journal of Toxicology- Cutaneous and Ocular Toxicology 23, no. 1 (2004): 19–28. http://dx.doi.org/10.1081/cus-120027484.

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4

Humpel, Christian. "Organotypic Brain Slice Cultures." Current Protocols in Immunology 123, no. 1 (October 12, 2018): e59. http://dx.doi.org/10.1002/cpim.59.

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Stahl, Katja, øivind Skare, and Reidun Torp. "Organotypic Cultures as a Model of Parkinson´s Disease. A Twist to an Old Model." Scientific World JOURNAL 9 (2009): 811–21. http://dx.doi.org/10.1100/tsw.2009.68.

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Organotypic cultures from the ventral mesencephalon (VM) are widely used to model Parkinson's disease (PD). In this method, neurotoxic compounds have traditionally been applied to the media to induce a uniform dopaminergic (DAergic) cell death in the tissue slices, regardless of the variation existing among slices. This study demonstrates a refinement of the toxic induction technique. We show that unilateral application of 6-hydroxydopamine (6-OHDA) at the tissue surface by means of a microelectrode causes a precisely localized cell death that closely resembles anin vivostereotactic model. This technique introduces an internal control that accounts for variation between slices and enables a precise quantification of the cell loss due to the toxin in use. We characterized organotypic VM cultures in terms of effects of 6-OHDA toxicity and number of DAergic neurons as judged by immunofluorescence and Western blots. Our findings illustrate that this new application technique greatly improves the representativeness of organotypic cultures as a model for PD.We characterized organotypic VM cultures in terms of effects of 6-OHDA toxicity and number of DAergic neurons as judged by immunofluorescence and Western blots. Our findings illustrate that this new application technique greatly improves the representativeness of organotypic cultures as a model for PD.

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Humpel, Christian. "Organotypic Brain Slices of ADULT Transgenic Mice: A Tool to Study Alzheimer’s Disease." Current Alzheimer Research 16, no. 2 (February 4, 2019): 172–81. http://dx.doi.org/10.2174/1567205016666181212153138.

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Transgenic mice have been extensively used to study the Alzheimer pathology. In order to reduce, refine and replace (3Rs) the number of animals, ex vivo cultures are used and optimized. Organotypic brain slices are the most potent ex vivo slice culture models, keeping the 3-dimensional structure of the brain and being closest to the in vivo situation. Organotypic brain slice cultures have been used for many decades but were mainly prepared from postnatal (day 8-10) old rats or mice. More recent work (including our lab) now aims to culture organotypic brain slices from adult mice including transgenic mice. Especially in Alzheimer´s disease research, brain slices from adult transgenic mice will be useful to study beta-amyloid plaques, tau pathology and glial activation. This review will summarize the studies using organotypic brain slice cultures from adult mice to mimic Alzheimer's disease and will highlight advantages and also pitfalls using this technique.

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Schuger, L., K. S. O'Shea, B. B. Nelson, and J. Varani. "Organotypic arrangement of mouse embryonic lung cells on a basement membrane extract: involvement of laminin." Development 110, no. 4 (December 1, 1990): 1091–99. http://dx.doi.org/10.1242/dev.110.4.1091.

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The behavior of embryonic murine lung cells on a basement membrane extract (Matrigel) was investigated. Single cell suspensions generated by trypsinization of lungs removed from day 12 embryos were plated on Matrigel and cultured for up to one week. The basement membrane extract was used as a gel, and as a wet or dried film. In all of these instances, organotypic arrangement of the embryonic lung cells was observed. This process consisted of cell aggregation, sorting, polarization and formation of a tridimensional organization resembling embryonic lung. The maximal degree of organotypic development was obtained by using a thick gel; minimal reorganization was observed using a dried film. A rabbit polyclonal serum to laminin inhibited organotypic pattern formation while normal rabbit serum did not. Culture of lung cells on laminin gels promoted epithelial cyst formation but poor mesenchymal organization. By studying the behavior of epithelial and/or mesenchymal enriched cell populations on Matrigel, it was concluded that organotypic pattern formation on Matrigel required the presence of both cell populations. Cultivation of dissociated lung cells on a gel consisting of a mixture of collagens type I and III (Vitrogen-100) produced only cell aggregation. Cultivation of lung cells on a thin film of Vitrogen-100 or on uncoated tissue culture plastic produced monolayers of mesenchymal cells alone. Cultivation of lung cells in suspension also failed to induce organotypic arrangement even at maximal cell densities. The present study strongly supports a role for the basement membrane in the organotypic rearrangement of embryonic lung cells and subsequent in vitro cyst formation and budding of the reestablished epithelium. This, in turn, reinforces the concept of the basement membrane as a major regulator of organogenesis.

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8

Rappoldt, Liam, Adrienne Weeks, Rodney Ouellete, Jeremy Roy, Catherine Taylor, Craig McCormick, Kathleen Attwood, and Inhwa Kim. "TMOD-26. ESTABLISHING A PATIENT-DERIVED, IN-VITRO ORGANOTYPIC SLICE CULTURE MODEL OF GBM." Neuro-Oncology 22, Supplement_2 (November 2020): ii233. http://dx.doi.org/10.1093/neuonc/noaa215.976.

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Abstract Glioblastoma Multiforme (GBM) is the most common primary malignant brain tumour. This tumour is universally fatal with a median survival of 15 months. Driving this pathology is an extremely heterogeneic tumour and complex tumour microenvironment. GBM research is primarily conducted using immortalized or primary cell lines due to their practicality and reproducibility. However, these cell lines do not effectively recapitulate the tumour microenvironment. Mouse models address these shortcomings but are laborious and expensive. We propose to utilize a patient derived organotypic culture model of GBM as an intermediary. We have utilized this model to test genetic manipulation via lentiviral transduction and the feasibility of utilizing this model to understand patient derived extracellular vesicles (EVs). We have sectioned and cultured patient derived organotypic models for 14 days without loss of viability. To determine if these organotypic cultures are amenable to lentiviral manipulation, tissue sections were transduced with far-red fluorescent lentivirus and efficiency determined by confocal laser scanning microscopy (CLSM) and flow cytometry (FC). To determine feasibility as a model for EVs, media obtained from patient-derived organotypic cultures was analyzed by western blot, nanoparticle tracking analysis (NTA), and nanoFlow Cytometry (nFC). In the future these EVs will be compared to those found in patient serum. The model of GBM has been lentivirally transduced to express a far-red fluorescent vector in approximately 15% of the slice, quantified by CLSM and FC. EV-sized particles positive for canonical EV markers have been identified in the media by NTA, nFC and western blot. Using lentiviral-mediated genetic engineering and emerging EV science, this organotypic slice culture models yields exciting utility in GBM research. The established organotypic slice culture model will likely be a valuable tool in the study of GBM biology and EV dynamics.

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9

Ozbun, Michelle A., and Craig Meyers. "Two Novel Promoters in the Upstream Regulatory Region of Human Papillomavirus Type 31b Are Negatively Regulated by Epithelial Differentiation." Journal of Virology 73, no. 4 (April 1, 1999): 3505–10. http://dx.doi.org/10.1128/jvi.73.4.3505-3510.1999.

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ABSTRACT Organotypic cultures support the stratification and differentiation of keratinocytes and the human papillomavirus (HPV) life cycle. We report transcription from four novel promoters in the HPV31b upstream regulatory region during the viral life cycle in organotypic cultures. Promoter initiation was not differentiation dependent; two promoters were down-regulated upon epithelial differentiation.

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10

Brulin, Bénédicte, John C. Nolan, Tecla Marangon, Milan Kovacevic, Mathias Chatelais, Pierre Meheust, Jérome Abadie, et al. "Evaluation of the Chemotherapy Drug Response Using Organotypic Cultures of Osteosarcoma Tumours from Mice Models and Canine Patients." Cancers 13, no. 19 (September 29, 2021): 4890. http://dx.doi.org/10.3390/cancers13194890.

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Improvements in the clinical outcome of osteosarcoma have plateaued in recent decades with poor translation between preclinical testing and clinical efficacy. Organotypic cultures retain key features of patient tumours, such as a myriad of cell types organized within an extracellular matrix, thereby presenting a more realistic and personalised screening of chemotherapeutic agents ex vivo. To test this concept for the first time in osteosarcoma, murine and canine osteosarcoma organotypic models were maintained for up to 21 days and in-depth analysis identified proportions of immune and stromal cells present at levels comparable to that reported in vivo in the literature. Cytotoxicity testing of a range of chemotherapeutic drugs (mafosfamide, cisplatin, methotrexate, etoposide, and doxorubicin) on murine organotypic culture ex vivo found limited response to treatment, with immune and stromal cells demonstrating enhanced survival over the global tumour cell population. Furthermore, significantly decreased sensitivity to a range of chemotherapeutics in 3D organotypic culture relative to 2D monolayer was observed, with subsequent investigation confirming reduced sensitivity in 3D than in 2D, even at equivalent levels of drug uptake. Finally, as proof of concept for the application of this model to personalised drug screening, chemotherapy testing with doxorubicin was performed on biopsies obtained from canine osteosarcoma patients. Together, this study highlights the importance of recapitulating the 3D tumour multicellular microenvironment to better predict drug response and provides evidence for the utility and possibilities of organotypic culture for enhanced preclinical selection and evaluation of chemotherapeutics targeting osteosarcoma.

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LI, JENNY J., STEVEN G. BRAMLET, EDWARD A. CARTER, and JOHN F. BURKE. "The Rat Lung Organotypic Culture." Journal of Trauma: Injury, Infection, and Critical Care 31, no. 2 (February 1991): 174–81. http://dx.doi.org/10.1097/00005373-199131020-00004.

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LI, JENNY J., STEVEN G. BRAMLET, EDWARD A. CARTER, and JOHN F. BURKE. "The Rat Lung Organotypic Culture." Journal of Trauma: Injury, Infection, and Critical Care 31, no. 2 (February 1991): 174–81. http://dx.doi.org/10.1097/00005373-199102000-00004.

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13

Gähwiler, B. H. "Organotypic cultures of neural tissue." Trends in Neurosciences 11, no. 11 (January 1988): 484–89. http://dx.doi.org/10.1016/0166-2236(88)90007-0.

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14

Kratochwil, K., F. Ekblom, N. Fusenig, G. Cunha, M. Darmon, and R. I. Freshney. "Organ development and organotypic culture." Cytotechnology 2, S3 (August 1989): 22–26. http://dx.doi.org/10.1007/bf02279719.

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15

Salerno, S., S. Morelli, E. Drioli, and L. De Bartolo. "Human Liver Organotypic Membrane Systems." Procedia Engineering 44 (2012): 456–58. http://dx.doi.org/10.1016/j.proeng.2012.08.448.

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16

Lyman, W. D., W. C. Hatch, E. Pousada, G. Stephney, W. K. Rashbaum, and K. M. Weidenheim. "Human fetal myelinated organotypic cultures." Brain Research 599, no. 1 (December 1992): 34–44. http://dx.doi.org/10.1016/0006-8993(92)90849-5.

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17

Sridhar, Adithya, Salvatore Simmini, Carla M. S. Ribeiro, Caroline Tapparel, Melvin M. Evers, Dasja Pajkrt, and Katja Wolthers. "A Perspective on Organoids for Virology Research." Viruses 12, no. 11 (November 23, 2020): 1341. http://dx.doi.org/10.3390/v12111341.

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Animal models and cell lines are invaluable for virology research and host–pathogen interaction studies. However, it is increasingly evident that these models are not sufficient to fully understand human viral diseases. With the advent of three-dimensional organotypic cultures, it is now possible to study viral infections in the human context. This perspective explores the potential of these organotypic cultures, also known as organoids, for virology research, antiviral testing, and shaping the virology landscape.

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Binder, Zev, Sarah Hyun Ji Kim, Pei-Hsun Wu, Anjil Giri, Gary Gallia, Carlos Pardo-Villamizar, and Denis Wirtz. "TMOD-12. THE ROLE OF INTEGRIN α V AND CD44 IN GBM MIGRATION USING HUMAN ORGANOTYPIC SLICES." Neuro-Oncology 21, Supplement_6 (November 2019): vi265. http://dx.doi.org/10.1093/neuonc/noz175.1111.

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Abstract Current model systems used for GBM research include traditional in vitro cell line-based assays and in vivo animal studies. In vitro model systems offer the advantages of being easy to use, relatively inexpensive, and fast growing. However, these models lack key elements of the pathology they are attempting to model, including the biochemical and biophysical microenvironment and three-dimensional structure inherent to human brain tissue. In vivo model systems address these limitations, but have restrictions of their own. Species differences may result in non-applicable results and animal experiments are often not designed like clinical trials. Evidence of the limitations of current GBM models is found in the disparity between basic research findings and successful new treatments for GBMs in the clinic. Here we present an alternative model system for the study of human GBM cell motility and invasion, which features advantages of both in vitro and in vivo model systems. Using human organotypic brain slices as scaffolding for tumor growth, we explored the dynamic process of GBM cell invasion within human brain tissue. To demonstrate the utility of the model system, we investigated the effects of depletion of integrin α V (ITGAV) and CD44 on GBM cell motility. These two cell-surface proteins have been identified to have key functions in GBM cell motility. However, knockdown of ITGAV had little effect on tumor cell motility in organotypics while CD44 knockdown significantly reduced cell movement. Finally, we compare motility results from cells in human brain slices to those from cells growing on standard Matrigel and in mouse brain organotypics. We found significant differences in motility depending on the substrate in which the cells were moving. Our findings highlight the physiologic characteristics of human brain organotypics and demonstrate the use of real-time imaging in the ex vivo system.

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Xiao, Suhong, Ying-Sheng Tang, Rehana A. Khan, Yonghua Zhang, Praveen Kusumanchi, Sally P. Stabler, Hiremagalur N. Jayaram, and Aśok C. Antony. "Influence of Physiologic Folate Deficiency on Human Papillomavirus Type 16 (HPV16)-harboring Human Keratinocytes in Vitro and in Vivo." Journal of Biological Chemistry 287, no. 15 (February 17, 2012): 12559–77. http://dx.doi.org/10.1074/jbc.m111.317040.

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Although HPV16 transforms infected epithelial tissues to cancer in the presence of several co-factors, there is insufficient molecular evidence that poor nutrition has any such role. Because physiological folate deficiency led to the intracellular homocysteinylation of heterogeneous nuclear ribonucleoprotein E1 (hnRNP-E1) and activated a nutrition-sensitive (homocysteine-responsive) posttranscriptional RNA operon that included interaction with HPV16 L2 mRNA, we investigated the functional consequences of folate deficiency on HPV16 in immortalized HPV16-harboring human (BC-1-Ep/SL) keratinocytes and HPV16-organotypic rafts. Although homocysteinylated hnRNP-E1 interacted with HPV16 L2 mRNA cis-element, it also specifically bound another HPV16 57-nucleotide poly(U)-rich cis-element in the early polyadenylation element (upstream of L2̂L1 genes) with greater affinity. Together, these interactions led to a profound reduction of both L1 and L2 mRNA and proteins without effects on HPV16 E6 and E7 in vitro, and in cultured keratinocyte monolayers and HPV16-low folate-organotypic rafts developed in physiological low folate medium. In addition, HPV16-low folate-organotypic rafts contained fewer HPV16 viral particles, a similar HPV16 DNA viral load, and a much greater extent of integration of HPV16 DNA into genomic DNA when compared with HPV16-high folate-organotypic rafts. Subcutaneous implantation of 18-day old HPV16-low folate-organotypic rafts into folate-replete immunodeficient mice transformed this benign keratinocyte-derived raft tissue into an aggressive HPV16-induced cancer within 12 weeks. Collectively, these studies establish a likely molecular linkage between poor folate nutrition and HPV16 and predict that nutritional folate and/or vitamin-B12 deficiency, which are both common worldwide, will alter the natural history of HPV16 infections and also warrant serious consideration as reversible co-factors in oncogenic transformation of HPV16-infected tissues to cancer.

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Hayman, Ian R., Rachel M. Temple, Cole K. Burgess, Mary Ferguson, Jason Liao, Craig Meyers, and Clare E. Sample. "New insight into Epstein-Barr Virus infection using models of stratified epithelium." PLOS Pathogens 19, no. 1 (January 11, 2023): e1011040. http://dx.doi.org/10.1371/journal.ppat.1011040.

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Epstein-Barr virus (EBV) is a ubiquitous human pathogen that is transmitted in saliva. EBV transits through the oral epithelium to infect B cells, where it establishes a life-long latent infection. Reinfection of the epithelium is believed to be mediated by virus shed from B cells, but whether a latent reservoir can exist in the epithelia is unknown. We previously developed an in vitro organotypic model of stratified epithelium where EBV can readily replicate within the suprabasal layers of the epithelium following apical infection mediated by virus-producing B cells. Given that infected epithelial cells and cell-free virus are observed in saliva, we examined the ability of both of these to mediate infection in organotypic cultures. Epithelial-derived cell-free virus was able to infect organotypic cultures from the apical surface, but showed enhanced infection of B cells. Conversely, B cell-derived virus exhibited enhanced infection of epithelial cells. While EBV has been detected in basal cells in oral hairy leukoplakia, it is unknown whether EBV can be seen in undifferentiated primary keratinocytes in the basal layer. Undifferentiated epithelial cells expressed proposed EBV receptors in monolayer and were susceptible to viral binding and entry. Integrins, and occasionally ephrin A2, were expressed in the basal layer of gingiva and tonsil derived organotypic cultures, but the known B-cell receptors HLAII and CD21 were not detected. Following infection with cell-free virus or virus-producing B cells at either the apical or basolateral surface of preformed organotypic cultures, abundant infection was detected in differentiated suprabasal cells while more limited but readily detectable infection was observed in the undifferentiated basal cells. Together, our data has provided new insight into EBV infection in stratified epithelium.

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Tsupykov, O., I. Lushnikova, Y. Nikandrova, K. Yatsenko, A. Ustymenko, V. Kyryk, G. Butenko, and G. Skybo. "A novel model of periventricular leukomalacia on mouse organotypic brain slice culture." Cell and Organ Transplantology 4, no. 2 (November 30, 2016): 188–93. http://dx.doi.org/10.22494/cot.v4i2.60.

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The creation of adequate in vitro and in vivo models of neural tissue injury is essential to assess the therapeutic effect of pharmacological agents and regenerative potential of various types of stem cells in diseases of the central nervous system. The aim of this work was to create a novel model of cerebral white matter lesions – periventricular leukomalacia (PVL) – on murine organotypic brain slice culture.Materials and methods. The PVL model was developed on cultured organotypic mice brain slices subjected to oxygen-glucose deprivation (OGD) followed by addition of endotoxin lipopolysaccharide (LPS) in the culture medium. To analyze the degree of tissue injury within PVL simulation, we used spectrophotometric method for estimation of cytosolic enzyme lactate dehydrogenase (LDH) in the culture medium and immunohistochemical analysis of the slices using antibodies to Rip, GFAP and Iba-1 protein markers of oligodendrocyte, astroglia and microglia, respectively.Results. It was shown that the combined effect of OGD and lipopolysaccharide resulted in a significant release of the cytosolic enzyme LDH in culture medium, decrease of Rip-immunoreactivity and a pronounced reactive astro- and microgliosis in murine organotypic brain slice culture.Conclusions. Our model of PVL developed on cultured organotypic mice brain slices is novel and promising tool to study pathogenic mechanisms of cerebral white matter lesions and ways of neuroprotection in this pathology, including pharmacological agents and transplantation of stem cells.

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22

Chu, Chih-Wen, and Lance A. Davidson. "Chambers for Culturing and Immobilizing Xenopus Embryos and Organotypic Explants for Live Imaging." Cold Spring Harbor Protocols 2022, no. 5 (October 19, 2021): pdb.prot107649. http://dx.doi.org/10.1101/pdb.prot107649.

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Live imaging of Xenopus embryos and organotypic explants can be challenging because of their large size and slippery nature. This protocol covers the preparation of special chambers for immobilizing Xenopus embryos and embryonic explants for live-cell and tissue imaging. The opaque nature of Xenopus embryonic tissues enables simple bright-field imaging techniques for tracking surface movements across large regions. Such surface imaging of embryos or organotypic explants can directly reveal cell behaviors, obviating the need for complex postprocessing commonly required to extract this data from 3D confocal or light-sheet observations of more transparent embryos. Furthermore, Xenopus embryos may be filled with light-absorbing pigment granules and light-scattering yolk platelets, but these limitations are offset by the utilitarian nature of Xenopus organotypic explants that expose and stabilize large embryonic cells in a nearly native context for high-resolution live-cell imaging. Additionally, whole embryos can be stabilized for long-term bright-field and confocal microscopy. Simple explants can be prepared using a single cell type, and organotypic explants can be prepared in which multiple tissue types are dissected while retaining native tissue–tissue interactions. These preparations enable both in-toto imaging of tissue dynamics and super-resolution imaging of protein dynamics within individual cells. We present detailed protocols for these methods together with references to applications.

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23

Egimendia, Ander, Susana Carregal-Romero, Iñaki Osorio-Querejeta, Daniel Padro, Jesús Ruiz-Cabello, David Otaegui, and Pedro Ramos-Cabrer. "Assessing the Potential of Molecular Imaging for Myelin Quantification in Organotypic Cultures." Pharmaceutics 13, no. 7 (June 28, 2021): 975. http://dx.doi.org/10.3390/pharmaceutics13070975.

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Ex vivo models for the noninvasive study of myelin-related diseases represent an essential tool to understand the mechanisms of diseases and develop therapies against them. Herein, we assessed the potential of multimodal imaging traceable myelin-targeting liposomes to quantify myelin in organotypic cultures. Methods: MRI testing was used to image mouse cerebellar tissue sections and organotypic cultures. Demyelination was induced by lysolecithin treatment. Myelin-targeting liposomes were synthetized and characterized, and their capacity to quantify myelin was tested by fluorescence imaging. Results: Imaging of freshly excised tissue sections ranging from 300 µm to 1 mm in thickness was achieved with good contrast between white (WM) and gray matter (GM) using T2w MRI. The typical loss of stiffness, WM structures, and thickness of organotypic cultures required the use of diffusion-weighted methods. Designed myelin-targeting liposomes allowed for semiquantitative detection by fluorescence, but the specificity for myelin was not consistent between assays due to the unspecific binding of liposomes. Conclusions: With respect to the sensitivity, imaging of brain tissue sections and organotypic cultures by MRI is feasible, and myelin-targeting nanosystems are a promising solution to quantify myelin ex vivo. With respect to specificity, fine tuning of the probe is required. Lipid-based systems may not be suitable for this goal, due to unspecific binding to tissues.

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Banerjee, N. Sanjib, Angel A. Rivera, Minghui Wang, Louise T. Chow, Thomas R. Broker, David T. Curiel, and Dirk M. Nettelbeck. "Analyses of melanoma-targeted oncolytic adenoviruses with tyrosinase enhancer/promoter-driven E1A, E4, or both in submerged cells and organotypic cultures." Molecular Cancer Therapeutics 3, no. 4 (April 1, 2004): 437–49. http://dx.doi.org/10.1158/1535-7163.437.3.4.

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Abstract We have generated novel conditionally replicative adenoviruses (CRAds) targeted to melanoma cells. In these adenoviruses, the E4 region (AdΔ24TyrE4) or both E1 and E4 regions (Ad2xTyr) were controlled by a synthetic tyrosinase enhancer/promoter (Tyr2E/P) specific for melanocytes. The properties of these CRAds were compared with wild-type adenovirus (Adwt) and our previous CRAd with a targeted E1A CRII mutation (AdTyrΔ24) in submerged cultures of melanoma cells and nonmelanoma control cells. We showed that AdΔ24TyrE4 had a cell type selectivity similar to AdTyrΔ24 but had a distinct block in viral reproduction in nonmelanoma cells and that Ad2xTyr had an augmented selectivity for melanoma cells. These viruses were additionally tested in organotypic cultures of melanoma cell lines, primary human keratinocytes (PHKs), or mixed cell populations. Unexpectedly, the CRAds exhibited somewhat different cell type selectivity profiles in these cultures relative to those observed in submerged cultures, demonstrating the importance of multiple assay systems. Specifically, AdTyrΔ24 and Ad2xTyr were selective for melanoma cells, whereas AdΔ24TyrE4 exhibited no selectivity, similar to Adwt. AdTyrΔ24 and Ad2xTyr were strongly attenuated in their ability to lyse PHKs in organotypic cultures. Furthermore, Ad2xTyr had a superior melanoma selectivity in organotypic cultures of cocultivated melanoma cells and PHKs. The enhanced selectivity for melanoma cells exhibited by Ad2xTyr provides a window of opportunity for therapeutic application. These studies also demonstrate that organotypic cultures derived from mixtures of tumor and normal cells represent a promising new model for analysis of CRAd specificity and toxicity.

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Kopanska, Katarzyna. "Cross-industrial applications of organotypic models." ALTEX 39, no. 1 (2022): 155–58. http://dx.doi.org/10.14573/altex.2112202.

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Takács, J., and F. Metzger. "Morphological study of organotypic cerebellar cultures." Acta Biologica Hungarica 53, no. 1-2 (March 2002): 187–204. http://dx.doi.org/10.1556/abiol.53.2002.1-2.18.

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Purcell, Wendy M., Christopher K. Atterwill, and Jinsheng Xu. "Cryopreservation of Organotypic Brain Spheroid Cultures." Alternatives to Laboratory Animals 31, no. 6 (November 2003): 563–73. http://dx.doi.org/10.1177/026119290303100605.

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Moldzio, Rudolf, Christina Piskernik, Khaled Radad, and Wolf-Dieter Rausch. "Rotenone Damages Striatal Organotypic Slice Culture." Annals of the New York Academy of Sciences 1148, no. 1 (December 2008): 530–35. http://dx.doi.org/10.1196/annals.1410.009.

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LYMAN, W. D., Y. KRESS, F. C. CHIU, C. S. RAINE, M. B. BBORNSTEIN, and A. RUVINSTEIN. "Human Fetal Neural Tissue Organotypic Cultures." Annals of the New York Academy of Sciences 546, no. 1 Molecular Bas (December 1988): 225–26. http://dx.doi.org/10.1111/j.1749-6632.1988.tb21647.x.

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SUGAI, Fuminobu, Yoichi YAMAMOTO, and Saburo SAKODA. "Organotypic spinal cord culture using mice." Folia Pharmacologica Japonica 124, no. 1 (2004): 19–23. http://dx.doi.org/10.1254/fpj.124.19.

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Koizumi, Amane, and Hiroshi Jouhou. "Organotypic culture of adult rodent retina." Neuroscience Research 65 (January 2009): S172. http://dx.doi.org/10.1016/j.neures.2009.09.902.

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Humpel, C. "Organotypic brain slice cultures: A review." Neuroscience 305 (October 2015): 86–98. http://dx.doi.org/10.1016/j.neuroscience.2015.07.086.

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Hoff, A., H. Hämmerle, and B. Schlosshauer. "Organotypic culture system of chicken retina." Brain Research Protocols 4, no. 3 (December 1999): 237–48. http://dx.doi.org/10.1016/s1385-299x(99)00024-0.

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Figiel, Sandy, Côme Pasqualin, Fanny Bery, Veronique Maupoil, Christophe Vandier, Marie Potier-Cartereau, Isabelle Domingo, et al. "Functional Organotypic Cultures of Prostate Tissues." American Journal of Pathology 189, no. 6 (June 2019): 1268–75. http://dx.doi.org/10.1016/j.ajpath.2019.02.017.

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Marx, Uwe. "Organotypic tissue culture for substance testing." Journal of Biotechnology 148, no. 1 (July 1, 2010): 1–2. http://dx.doi.org/10.1016/j.jbiotec.2010.06.001.

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Kloth, Sabine, Elfriede Eckert, Stefan J. Klein, Jan Monzer, Christiane Wanke, and Will W. Minuth. "Gastric epithelium under organotypic perfusion culture." In Vitro Cellular & Developmental Biology - Animal 34, no. 7 (August 1998): 515–17. http://dx.doi.org/10.1007/s11626-998-0107-9.

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Avner, Ellis D., Nicholas P. Piesco, William E. Sweeney, and Demetrius Ellis. "Renal epithelial development in organotypic culture." Pediatric Nephrology 2, no. 1 (March 1988): 92–99. http://dx.doi.org/10.1007/bf00870387.

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Oda, Dolphine P., Lydia Eng, Christopher E. Savard, Mike Newcomer, W. Geoffrey Haigh, and Sum P. Lee. "Organotypic Culture of Human Gallbladder Epithelium." Experimental and Molecular Pathology 63, no. 1 (August 1995): 16–22. http://dx.doi.org/10.1006/exmp.1995.1026.

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Gähwiler, B. H. "Organotypic slice cultures of neural tissue." Neuroscience Research Supplements 16 (January 1991): XIV. http://dx.doi.org/10.1016/0921-8696(91)90634-y.

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Reiner, Peter B., Bernd Heimrich, Flavio Keller, and Helmut L. Haas. "Organotypic culture of central histamine neurons." Brain Research 442, no. 1 (February 1988): 166–70. http://dx.doi.org/10.1016/0006-8993(88)91446-1.

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Schweiger, Pawel J., and Kim B. Jensen. "Modeling human disease using organotypic cultures." Current Opinion in Cell Biology 43 (December 2016): 22–29. http://dx.doi.org/10.1016/j.ceb.2016.07.003.

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Caesar, P. A., S. J. Wilson, C. S. Normand, and A. D. Postle. "A comparison of the specificity of phosphatidylcholine synthesis by human fetal lung maintained in either organ or organotypic culture." Biochemical Journal 253, no. 2 (July 15, 1988): 451–57. http://dx.doi.org/10.1042/bj2530451.

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Human fetal lung (14-18 weeks gestation) was maintained in either organ or organotypic culture. By 4 days in organ culture or 14 days in organotypic culture, epithelial cells within both culture systems exhibited well-developed apical microvilli and possessed numerous intracellular lamellar bodies characteristic of surfactant phospholipid stores. However, analysis of the pattern of synthesis of individual molecular species of phosphatidylcholine by [14C]choline incorporation and reversed-phase h.p.l.c. showed that this apparent maturation was not paralleled by an increased synthesis of the dipalmitoyl species in either culture system. By contrast, the fractional synthesis of dipalmitoyl phosphatidylcholine, expressed as a percentage of total [14C]choline incorporation, decreased with time in both organ and organotypic culture. Moreover, these fractions were not significantly different from those measured in parallel monolayer cultures of mixed human fetal lung cells that displayed mainly fibroblast morphology. These results suggest that the synthesis pattern of phosphatidylcholine species by lung cells in culture is determined principally by their incubation conditions and not by their state of apparent maturation.

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Weitz, Jonathan, Tatiana Hurtado de Mendoza, Herve Tiriac, Joel Baumgartner, Kaitlyn Kelly, Jula Veerapong, and Andrew Lowy. "Abstract 289: A novel ex-vivo organotypic culture platform for functional interrogation of human appendiceal neoplasms." Cancer Research 82, no. 12_Supplement (June 15, 2022): 289. http://dx.doi.org/10.1158/1538-7445.am2022-289.

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Abstract Appendiceal neoplasms are rare and often clinically present with peritoneal metastasis. While surgical tumor resection is effective for some types of primary appendiceal cancers, patients with metastatic disease have poor prognostic outcomes. Models to study appendix cancer biology are limited, given that 1) no mouse models exist and 2) reliable in vitro models are unavailable. As such, we have developed an ex-vivo organotypic slice model to examine cellular interactions between tumor cells and their local microenvironment. Tumor specimens from human appendiceal cancer patients were cut using a vibratome to make 200 μm organotypic slices. Slices were cultured on transwell inserts and tested for changes in morphological, cellular and functional characteristics over a seven-day period. Organotypic slices maintained their cellular composition in regard to the proportion of epithelial, immune cells and fibroblasts. Live cell [Ca2+]i imaging of long term cultured slices confirmed that immune cells remain functionally active when stimulated with extracellular ATP. Lasty, using tumor biopsies from human donors, we have identified a diverse immunological profile of appendiceal tumors not previously identified. Our study illustrates a novel approach for studying the pathophysiology of appendiceal cancer, a notoriously difficult disease to model. Citation Format: Jonathan Weitz, Tatiana Hurtado de Mendoza, Herve Tiriac, Joel Baumgartner, Kaitlyn Kelly, Jula Veerapong, Andrew Lowy. A novel ex-vivo organotypic culture platform for functional interrogation of human appendiceal neoplasms [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 289.

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Chang, Tim C., Andrei M. Mikheev, Wilson Huynh, Raymond J. Monnat, Robert C. Rostomily, and Albert Folch. "Parallel microfluidic chemosensitivity testing on individual slice cultures." Lab Chip 14, no. 23 (2014): 4540–51. http://dx.doi.org/10.1039/c4lc00642a.

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Short, Ben. "Time (and PPARβ/δ) heals all wounds." Journal of Cell Biology 184, no. 6 (March 23, 2009): 767. http://dx.doi.org/10.1083/jcb.1846if.

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Hedberg, Jesper J., Annette Hansson, Jan A. Nilsson, Jan-Olov Höög, and Roland C. Grafström. "Uniform Expression of Alcohol Dehydrogenase 3 in Epithelia Regenerated with Cultured Normal, Immortalised and Malignant Human Oral Keratinocytes." Alternatives to Laboratory Animals 29, no. 3 (May 2001): 325–33. http://dx.doi.org/10.1177/026119290102900308.

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The human oral epithelium is a target for damage from the inhalation of formaldehyde. However, most experimental studies on this chemical have relied on laboratory animals that are obligatory nose breathers, including rats and mice. Therefore, in vitro model systems that mimic the structure of the human oral epithelium and which retain normal tissue-specific metabolic competence are desirable. Based on the established role of alcohol dehydrogenase 3 (ADH3), also known as glutathione-dependent formaldehyde dehydrogenase, as the primary enzyme catalysing the detoxification of formaldehyde, the aim of this study was to investigate the expression of ADH3 in organotypic epithelia regenerated with normal (NOK), immortalised (SVpgC2a) and malignant (SqCC/Y1) human oral keratinocytes. Organotypic epithelia, usually consisting of 5–10 cell layers, were produced at the air–liquid interface of collagen gels containing human oral fibroblasts, after culture for 10 days in a standardised serum-free medium. Immunochemical staining demonstrated uniform expression of ADH3 in these organotypic epithelia, as well as in the epithelial cells of oral tissue. The specificity of the ADH3 antiserum was ascertained from the complete neutralisation of the immunochemical reaction with purified ADH3 protein. Assessment of the staining intensities indicated that the expression levels were similar among the regenerated epithelia. Furthermore, the regenerated epithelia showed similar ADH3 expression to the epithelium in oral tissue. Therefore, a tissue-like expression pattern for ADH3 can be generated from the culture of various oral keratinocyte lines in an organotypic state. Similar expression levels among the various cell lines indicate the preservation of ADH3 during malignant transformation, and therefore that NOK, SVpgC2a and SqCC/Y1 represent functional models for in vitro studies of formaldehyde metabolism in human oral mucosa.

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Spahr-Schopfer, Isabelle, Lazlo Vutskits, Nicholas Toni, Pierre-Alain Buchs, Lorena Parisi, and Dominique Muller. "Differential Neurotoxic Effects of Propofol on Dissociated Cortical Cells and Organotypic Hippocampal Cultures." Anesthesiology 92, no. 5 (May 1, 2000): 1408–17. http://dx.doi.org/10.1097/00000542-200005000-00032.

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Background Propofol is a widely used anesthetic agent for adults and children. Although extensive clinical use has demonstrated its safety, neurologic dysfunctions have been described after the use of this agent. A recent study on a model of aggregating cell cultures reported that propofol might cause irreversible lesions of gamma-aminobutyric acid-mediated (GABAergic) neurons when administered at a critical phase of brain development. We investigated this issue by comparing the effects of long-term propofol treatment on two models of brain cultures: dissociated neonatal cortical cell cultures and organotypic slice cultures. Methods Survival of GABAergic neurons in dissociated cultures of newborn rat cortex (postnatal age, 1 day) treated for 3 days with different concentrations of propofol was assessed using histologic and cytochemical methods. For hippocampal organotypic slice cultures (postnatal age, 1 and 7 days), cell survival was assessed by measuring functional and morphologic parameters: extracellular and intracellular electrophysiology, propidium staining of dying cells, and light and electron microscopy. Results In dissociated neonatal cell cultures, propofol induced dose-dependent lesions of GABAergic neurons and of glial cells. In contrast, no evidence for neurotoxic effects of propofol were found after long-term treatment of organotypic slice cultures. Excitatory transmission was not affected by propofol, and inhibitory transmission was still functional. Histologic preparations showed no evidence for cell degeneration or death. Conclusion Although long-term applications of propofol to dissociated cortical cell cultures produced degeneration and death of GABAergic neurons and glial cells, no such lesions were found when using a model of postnatal organotypic slice cultures. This conclusion is based on both functional and morphologic tests.

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GIANINAZZI, C., M. SCHILD, N. MÜLLER, S. L. LEIB, F. SIMON, S. NUÑEZ, P. JOSS, and B. GOTTSTEIN. "Organotypic slice cultures from rat brain tissue: a new approach forNaegleria fowleriCNS infectionin vitro." Parasitology 132, no. 6 (September 13, 2005): 797–804. http://dx.doi.org/10.1017/s0031182005008619.

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The free-living amoebaNaegleria fowleriis the aetiological agent of primary amoebic meningoencephalitis (PAM), a disease leading to death in the vast majority of cases. In patients suffering from PAM, and in corresponding animal models, the brain undergoes a massive inflammatory response, followed by haemorrhage and severe tissue necrosis. Both,in vivoandin vitromodels are currently being used to study PAM infection. However, animal models may pose ethical issues, are dependent upon availability of specific infrastructural facilities, and are time-consuming and costly. Conversely, cell cultures lack the complex organ-specific morphology foundin vivo, and thus, findings obtainedin vitrodo not necessarily reflect the situationin vivo. The present study reports infection of organotypic slice cultures from rat brain withN. fowleriand compares the findings in this culture system within vivoinfection in a rat model of PAM, that proved complementary to that of mice. We found that brain morphology, as presentin vivo, is well retained in organotypic slice cultures, and that infection time-course including tissue damage parallels the observationsin vivoin the rat. Therefore, organotypic slice cultures from rat brain offer a newin vitroapproach to studyN. fowleriinfection in the context of PAM.

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Joshi, Nidhi, Dongli Liu, Kristie-Ann Dickson, Deborah J. Marsh, Caroline E. Ford, and Martina H. Stenzel. "An organotypic model of high-grade serous ovarian cancer to test the anti-metastatic potential of ROR2 targeted Polyion complex nanoparticles." Journal of Materials Chemistry B 9, no. 44 (2021): 9123–35. http://dx.doi.org/10.1039/d1tb01837j.

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Pantazopoulou, Vasiliki, Tracy Berg, and Alexander Pietras. "TAMI-71. EFFECTS OF THE TREATED MICROENVIRONMENT ON GLIOMA CELL PROPERTIES IN AN ORGANOTYPIC BRAIN SLICE MODEL." Neuro-Oncology 23, Supplement_6 (November 2, 2021): vi213. http://dx.doi.org/10.1093/neuonc/noab196.853.

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Abstract Glioblastoma is the most aggressive primary brain tumor. Despite treatment all patients invariably recur. Treatment resistance is attributed to the presence of glioma stem-like cells. Initially thought to be a distinct and static cell population, it is becoming increasingly clear that the glioma stem-like cell phenotype represents one of many cellular states and that glioma cells show plasticity between stem-like and non-stem like states. These plastic cell states are affected by the tumor microenvironment. In our lab we have shown that irradiated and hypoxic astrocytes increase the stem-like cell properties of glioma cells. In this study, we aim to evaluate how the treated microenvironment alters glioma cell properties and use ex vivo organotypic brain slices generated from tumor bearing and tumor naïve mice to assess all aspects of the microenvironment. We first characterized organotypic brain slices cultured in different oxygen tensions. We saw that tumor-bearing slices survive for at least 14 days in culture at 21%, 5% or 1% oxygen tension (O2). Tumor cells were more viable in all culture conditions and timepoints compared to non-tumor cells. Moreover, we found that astrocytes seem to be attracted to tumor areas in both 5% and 1% O2 cultures. We then used the organotypic glioma slice culture system to address how preconditioning the microenvironment using radiation or temozolomide affects the properties of glioma cells that are seeded in these pretreated, tumor naïve slices. We saw that fluorescently labelled glioma cells seeded in treated slices can be isolated after two days of culture in the slices and can be used for downstream analyses, such as temozolomide or radiation treatment and colony formation. This study will elucidate the effect of the treated microenvironment on glioma cell properties by using the medium throughput method of organotypic slice cultures.

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