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Stem Cell Reports. 2016 May 10;6(5):717-728. doi: 10.1016/j.stemcr.2016.04.001. Epub 2016 Apr 28.

Optimized Approaches for Generation of Integration-free iPSCs from Human Urine-Derived Cells with Small Molecules and Autologous Feeder.

Author information

1
Department of Hematology, ZhuJiang Hospital of Southern Medical University, Guangzhou, Guangdong 510280, China.
2
CAS Key Laboratory of Regenerative Biology, South China Institute for Stem Cell Biology and Regenerative Medicine, Guangzhou Institutes of Biomedicine and Health, Chinese Academy of Sciences, Guangdong Provincial Key Laboratory of Stem Cell and Regenerative Medicine, Guangzhou, Guangdong 510530, China.
3
Department of Hematology, ZhuJiang Hospital of Southern Medical University, Guangzhou, Guangdong 510280, China. Electronic address: liyuhua2011gz@163.com.
4
Department of Hematology, ZhuJiang Hospital of Southern Medical University, Guangzhou, Guangdong 510280, China. Electronic address: jimzhua@163.com.
5
CAS Key Laboratory of Regenerative Biology, South China Institute for Stem Cell Biology and Regenerative Medicine, Guangzhou Institutes of Biomedicine and Health, Chinese Academy of Sciences, Guangdong Provincial Key Laboratory of Stem Cell and Regenerative Medicine, Guangzhou, Guangdong 510530, China. Electronic address: pan_guangjin@gibh.ac.cn.

Abstract

Generation of induced pluripotent stem cells (iPSCs) from human urine-derived cells (hUCs) provides a convenient and non-invasive way to obtain patient-specific iPSCs. However, many isolated hUCs exhibit very poor proliferation and are difficult to reprogram. In this study, we optimized reprogramming approaches for hUCs with very poor proliferation. We report here that a compound cocktail containing cyclic pifithrin-a (a P53 inhibitor), A-83-01, CHIR99021, thiazovivin, NaB, and PD0325901 significantly improves the reprogramming efficiency (170-fold more) for hUCs. In addition, we showed that replacement of Matrigel with autologous hUC feeders can overcome the reprogramming failure due to the massive cell death that occurs during delivery of reprogramming factors. In summary, we describe improved approaches to enable iPSC generation from hUCs that were otherwise difficult to reprogram, a valuable asset for banking patient-specific iPSCs.

KEYWORDS:

CPFT-a; P53; autologous UC feeders; hUCs; iPSCs; small molecules

PMID:
27132887
PMCID:
PMC4939659
DOI:
10.1016/j.stemcr.2016.04.001
[Indexed for MEDLINE]
Free PMC Article

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