Conditional reprogramming and long-term expansion of normal and tumor cells from human biospecimens
| Autor: | Christopher Albanese, Thomas N. Darling, Seema Agarwal, Anatoly Dritschilo, Nancy Palechor-Ceron, Rajesh L. Thangapazham, Sujata Choudhury, Scott H. Randell, Ewa Krawczyk, Richard Schlegel, Hang Yuan, Tung-Wei Hou, Jie Lu, Frank A. Suprynowicz, Yun-Ling Zheng, Praathibha Sripadhan, Bhaskar Kallakury, Vera Simic, Xuefeng Liu, Chen Chen, Olga A. Timofeeva, Aleksandra Dakic, Dean G. Tang |
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| Rok vydání: | 2017 |
| Předmět: |
0301 basic medicine
Pyridines Biology Regenerative medicine General Biochemistry Genetics and Molecular Biology Article 03 medical and health sciences Tissue culture Mice Neoplasms Animals Humans Protein Kinase Inhibitors rho-Associated Kinases Drug discovery Feeder Cells Fibroblasts Cellular Reprogramming Amides In vitro Coculture Techniques Cell biology Rats Gene expression profiling 030104 developmental biology Cell Transformation Neoplastic Cell culture Immunology Cryopreserved Tissue Reprogramming |
| Zdroj: | Nature protocols. 12(2) |
| ISSN: | 1750-2799 |
| Popis: | Historically, it has been difficult to propagate cells in vitro that are derived directly from human tumors or healthy tissue. However, in vitro preclinical models are essential tools for both the study of basic cancer biology and the promotion of translational research, including drug discovery and drug target identification. This protocol describes conditional reprogramming (CR), which involves coculture of irradiated mouse fibroblast feeder cells with normal and tumor human epithelial cells in the presence of a Rho kinase inhibitor (Y-27632). CR cells can be used for various applications, including regenerative medicine, drug sensitivity testing, gene expression profiling and xenograft studies. The method requires a pathologist to differentiate healthy tissue from tumor tissue, and basic tissue culture skills. The protocol can be used with cells derived from both fresh and cryopreserved tissue samples. As approximately 1 million cells can be generated in 7 d, the technique is directly applicable to diagnostic and predictive medicine. Moreover, the epithelial cells can be propagated indefinitely in vitro, yet retain the capacity to become fully differentiated when placed into conditions that mimic their natural environment. |
| Databáze: | OpenAIRE |
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