Conditional reprogramming: next generation cell culture

Xiaoxiao Wu; Shengpeng Wang; Mingxing Li; Jing Li; Jing Shen; Yueshui Zhao; Jun Pang; Qinglian Wen; Meijuan Chen; Bin Wei; Parham Jabbarzadeh Kaboli; Fukuan Du; Qijie Zhao; Chi Hin Cho; Yitao Wang; Zhangang Xiao; Xu Wu

doi:10.1016/j.apsb.2020.01.011

Conditional reprogramming: next generation cell culture

Acta Pharm Sin B. 2020 Aug;10(8):1360-1381. doi: 10.1016/j.apsb.2020.01.011. Epub 2020 Jan 26.

Authors

Xiaoxiao Wu^{1

2}, Shengpeng Wang³, Mingxing Li^{1

2}, Jing Li⁴, Jing Shen^{1

2}, Yueshui Zhao^{1

2}, Jun Pang⁵, Qinglian Wen⁶, Meijuan Chen^{1

2}, Bin Wei⁷, Parham Jabbarzadeh Kaboli^{1

2}, Fukuan Du^{1

2}, Qijie Zhao^{1

2}, Chi Hin Cho^{1

2

8}, Yitao Wang³, Zhangang Xiao^{1

2}, Xu Wu^{1

2}

Affiliations

¹ Laboratory of Molecular Pharmacology, Department of Pharmacology, School of Pharmacy, Southwest Medical University, Luzhou 646000, China.
² South Sichuan Institute of Translational Medicine, Luzhou 646000, China.
³ State Key Laboratory of Quality Research in Chinese Medicine, Institute of Chinese Medical Sciences, University of Macau, Macao, China.
⁴ Department of Oncology and Hematology, Hospital (T.C.M) Affiliated to Southwest Medical University, Luzhou 646000, China.
⁵ Center of Radiation Oncology, Hospital (T.C.M) Affiliated to Southwest Medical University, Luzhou 646000, China.
⁶ Department of Oncology, the Affiliated Hospital of Southwest Medical University, Southwest Medical University, Luzhou 646000, China.
⁷ College of Pharmaceutical Science & Collaborative Innovation Center of Yangtze River Delta Region Green Pharmaceuticals, Zhejiang University of Technology, Hangzhou 310014, China.
⁸ School of Biomedical Sciences, Faculty of Medicine, the Chinese University of Hong Kong, Hong Kong, China.

Abstract

Long-term primary culture of mammalian cells has been always difficult due to unavoidable senescence. Conventional methods for generating immortalized cell lines usually require manipulation of genome which leads to change of important biological and genetic characteristics. Recently, conditional reprogramming (CR) emerges as a novel next generation tool for long-term culture of primary epithelium cells derived from almost all origins without alteration of genetic background of primary cells. CR co-cultures primary cells with inactivated mouse 3T3-J2 fibroblasts in the presence of RHO-related protein kinase (ROCK) inhibitor Y-27632, enabling primary cells to acquire stem-like characteristics while retain their ability to fully differentiate. With only a few years' development, CR shows broad prospects in applications in varied areas including disease modeling, regenerative medicine, drug evaluation, drug discovery as well as precision medicine. This review is thus to comprehensively summarize and assess current progress in understanding mechanism of CR and its wide applications, highlighting the value of CR in both basic and translational researches and discussing the challenges faced with CR.

Keywords: 3T3-J2 fibroblast; AACR, American Association for Cancer Research; ACC, adenoid cystic carcinoma; AR, androgen receptor; CFTR, cystic fibrosis transmembrane conductance regulators; CR, conditional reprogramming; CYPs, cytochrome P450 enzymes; Conditional reprogramming; DCIS, ductal carcinoma in situ; ECM, extracellular matrix; ESC, embryonic stem cell; HCMI, human cancer model initiatives; HGF, hepatocyte growth factor; HNE, human nasal epithelial; HPV, human papillomaviruses; ICD, intracellular domain; LECs, limbal epithelial cells; NCI, National Cancer Institute; NGFR, nerve growth factor receptor; NSCLC, non-small cell lung cancer; NSG, NOD/SCID/gamma; PDAC, pancreatic ductal adenocarcinoma; PDX, patient derived xenograft; PP2A, protein phosphatase 2A; RB, retinoblastoma-associated protein; ROCK; ROCK, Rho kinase; SV40, simian virus 40 large tumor antigen; Senescence; UVB, ultraviolet radiation b; Y-27632; dECM, decellularized extracellular matrix; hASC, human adipose stem cells; hTERT, human telomerase reverse transcriptase; iPSCs, induction of pluripotent stem cells; ΔNP63α, N-terminal truncated form of P63α.

Publication types

Review