title: Optimized Approaches for Generation of Integration-free iPSCs from Human Urine-Derived Cells with Small Molecules and Autologous Feeder.
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.
authors.affiliation: Department of Hematology, ZhuJiang Hospital of Southern Medical University, Guangzhou, Guangdong 510280, China.
authors.forename: Di
authors.initials: D
authors.lastname: Li
authors.affiliation: 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.
authors.forename: Linli
authors.initials: L
authors.lastname: Wang
authors.affiliation: 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.
authors.forename: Jundi
authors.initials: J
authors.lastname: Hou
authors.affiliation: Department of Hematology, ZhuJiang Hospital of Southern Medical University, Guangzhou, Guangdong 510280, China.
authors.forename: Qun
authors.initials: Q
authors.lastname: Shen
authors.affiliation: 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.
authors.forename: Qianyu
authors.initials: Q
authors.lastname: Chen
authors.affiliation: 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.
authors.forename: Xiaoshan
authors.initials: X
authors.lastname: Wang
authors.affiliation: 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.
authors.forename: Juan
authors.initials: J
authors.lastname: Du
authors.affiliation: 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.
authors.forename: Xiujuan
authors.initials: X
authors.lastname: Cai
authors.affiliation: 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.
authors.forename: Yongli
authors.initials: Y
authors.lastname: Shan
authors.affiliation: 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.
authors.forename: Tian
authors.initials: T
authors.lastname: Zhang
authors.affiliation: 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.
authors.forename: Tiancheng
authors.initials: T
authors.lastname: Zhou
authors.affiliation: 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.
authors.forename: Xi
authors.initials: X
authors.lastname: Shi
authors.affiliation: Department of Hematology, ZhuJiang Hospital of Southern Medical University, Guangzhou, Guangdong 510280, China. Electronic address: liyuhua2011gz@163.com.
authors.forename: Yuhua
authors.initials: Y
authors.lastname: Li
authors.affiliation: Department of Hematology, ZhuJiang Hospital of Southern Medical University, Guangzhou, Guangdong 510280, China. Electronic address: jimzhua@163.com.
authors.forename: Hua
authors.initials: H
authors.lastname: Zhang
authors.affiliation: 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.
authors.forename: Guangjin
authors.initials: G
authors.lastname: Pan
mesh_terms: D001549:Benzamides / Q000494:pharmacology; D052160:Benzothiazoles / Q000494:pharmacology; D002454:Cell Differentiation / Q000187:drug effects*; D049109:Cell Proliferation / Q000187:drug effects*; D065150:Cellular Reprogramming / Q000187:drug effects*; D004159:Diphenylamine / Q000031:analogs & derivatives / Q000494:pharmacology; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000166:cytology / Q000187:drug effects*; D011720:Pyrazoles / Q000494:pharmacology; D011725:Pyridines / Q000494:pharmacology; D011743:Pyrimidines / Q000494:pharmacology; D013844:Thiazoles / Q000494:pharmacology; D013882:Thiosemicarbazones / Q000494:pharmacology; D014050:Toluene / Q000031:analogs & derivatives / Q000494:pharmacology; D014556:Urine / Q000166:cytology
chemical_list: C507011:A-83-01; D001549:Benzamides; D052160:Benzothiazoles; C473711:Chir 99021; D011720:Pyrazoles; D011725:Pyridines; D011743:Pyrimidines; D013844:Thiazoles; D013882:Thiosemicarbazones; C545214:thiazovivin; D014050:Toluene; C506614:mirdametinib; D004159:Diphenylamine; C121565:pifithrin
keywords: CPFT-a; P53; autologous UC feeders; hUCs; iPSCs; small molecules