Conditional reprogramming and long-term expansion of normal and tumor cells from human biospecimens

Developers

Xuefeng Liu, Ewa Krawczyk, Seema Agarwal, Richard Schlegel, etc.

Description of the technology

The cells that are derived directly from human tumors or healthy tissue are difficult to propagate in vitro. The establishment of tumor cell lines is hindered by the low rate of success (1–10%, depending on the tissue of origin and state of disease progression) and high heterogeneity of primary tumors, which can not be reproduced in culture in its entirety. 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 technology is used for 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). The method requires a pathologist to differentiate healthy tissue from tumor tissue, and basic tissue culture skills. The protocol according to the technology 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.

Practical application

The technology is applicable to establish primary epithelial cell cultures in vitro from healthy human tissue and human cancer samples. Reprogrammed cells can be used for various applications, including regenerative medicine, drug sensitivity testing, gene expression profiling and xenograft studies. The technology can be valuable for development of patient-derived xenografts and organoid cultures. In addition, it has exciting potential as a method to be used as a complementary platform for basic, translational and clinical research.

Laboratories

• Department of Pathology, Georgetown University Medical Center, Washington (USA)
• Center for Cell Reprogramming, Georgetown University Medical Center Washington (USA)
• Department of Oncology, Georgetown University Medical Center, Washington (USA)

Links

Publications

• Liu, X. et al. «Conditional reprogramming and long-term expansion of normal and tumor cells from human biospecimens." 12 Nature Protocols (2017): 439–451.
• Liu, X. et al. «ROCK inhibitor and feeder cells induce the conditional reprogramming of epithelial cells." 180 Am. J. Pathol. (2012): 599–607.
• Fulcher, M.L. & Randell, S.H. «Human nasal and tracheo-bronchial respiratory cell culture." 945 Methods Mol. Biol. (2013): 109–121.