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

Xuefeng Liu, Ewa Krawczyk, Frank A. Suprynowicz, Nancy Palechor-Ceron, Hang Yuan, Aleksandra Dakic, Vera Simic, Yun Ling Zheng, Praathibha Sripadhan, Chen Chen, Jie Lu, Tung Wei Hou, Sujata Choudhury, Bhaskar Kallakury, Dean Tang, Thomas Darling, Rajesh Thangapazham, Olga Timofeeva, Anatoly Dritschilo, Scott H. Randell & 3 others Christopher Albanese, Seema Agarwal, Richard Schlegel

Research output: Research - peer-reviewArticle

  • 3 Citations

Abstract

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.

LanguageEnglish (US)
Pages439-451
Number of pages13
JournalNature Protocols
Volume12
Issue number2
DOIs
StatePublished - Feb 1 2017

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Tumors
Cells
Tissue
Neoplasms
In Vitro Techniques
Pharmaceutical Preparations
Epithelial Cells
Feeder Cells
rho-Associated Kinases
Tissue culture
Fibroblasts
Heterografts
Gene expression
Medicine
Testing
Regenerative Medicine
Y 27632
Drug Discovery
Translational Medical Research
Gene Expression Profiling

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)

Cite this

Liu, X., Krawczyk, E., Suprynowicz, F. A., Palechor-Ceron, N., Yuan, H., Dakic, A., ... Schlegel, R. (2017). Conditional reprogramming and long-term expansion of normal and tumor cells from human biospecimens. Nature Protocols, 12(2), 439-451. DOI: 10.1038/nprot.2016.174

Conditional reprogramming and long-term expansion of normal and tumor cells from human biospecimens. / Liu, Xuefeng; Krawczyk, Ewa; Suprynowicz, Frank A.; Palechor-Ceron, Nancy; Yuan, Hang; Dakic, Aleksandra; Simic, Vera; Zheng, Yun Ling; Sripadhan, Praathibha; Chen, Chen; Lu, Jie; Hou, Tung Wei; Choudhury, Sujata; Kallakury, Bhaskar; Tang, Dean; Darling, Thomas; Thangapazham, Rajesh; Timofeeva, Olga; Dritschilo, Anatoly; Randell, Scott H.; Albanese, Christopher; Agarwal, Seema; Schlegel, Richard.

In: Nature Protocols, Vol. 12, No. 2, 01.02.2017, p. 439-451.

Research output: Research - peer-reviewArticle

Liu, X, Krawczyk, E, Suprynowicz, FA, Palechor-Ceron, N, Yuan, H, Dakic, A, Simic, V, Zheng, YL, Sripadhan, P, Chen, C, Lu, J, Hou, TW, Choudhury, S, Kallakury, B, Tang, D, Darling, T, Thangapazham, R, Timofeeva, O, Dritschilo, A, Randell, SH, Albanese, C, Agarwal, S & Schlegel, R 2017, 'Conditional reprogramming and long-term expansion of normal and tumor cells from human biospecimens' Nature Protocols, vol 12, no. 2, pp. 439-451. DOI: 10.1038/nprot.2016.174
Liu X, Krawczyk E, Suprynowicz FA, Palechor-Ceron N, Yuan H, Dakic A et al. Conditional reprogramming and long-term expansion of normal and tumor cells from human biospecimens. Nature Protocols. 2017 Feb 1;12(2):439-451. Available from, DOI: 10.1038/nprot.2016.174
Liu, Xuefeng ; Krawczyk, Ewa ; Suprynowicz, Frank A. ; Palechor-Ceron, Nancy ; Yuan, Hang ; Dakic, Aleksandra ; Simic, Vera ; Zheng, Yun Ling ; Sripadhan, Praathibha ; Chen, Chen ; Lu, Jie ; Hou, Tung Wei ; Choudhury, Sujata ; Kallakury, Bhaskar ; Tang, Dean ; Darling, Thomas ; Thangapazham, Rajesh ; Timofeeva, Olga ; Dritschilo, Anatoly ; Randell, Scott H. ; Albanese, Christopher ; Agarwal, Seema ; Schlegel, Richard. / Conditional reprogramming and long-term expansion of normal and tumor cells from human biospecimens. In: Nature Protocols. 2017 ; Vol. 12, No. 2. pp. 439-451
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