机构:[1]Stem Cell Engineering Laboratory of Yunnan Province, Kunming General Hospital of Chengdu Military Command, Kunming, 650032, P. R. China[2]Neurology Department, The First Affiliated Hospital, Kunming Medical University, Kunming, P. R. China内科科室外科科室神经内科泌尿外科昆明医科大学附属第一医院[3]Center for Experiment, Yunnan Agricultural University, Kunming, 650201, P. R. China[4]Department of Neurology, The First Affiliated Hospital, Sun Yat-sen University, Guangzhou, P. R. China外科科室神经科泌尿外科中山大学附属第一医院
Multipotent stem cells have potential therapeutic roles in the treatment of Duchenne muscular dystrophy (DMD). However, the limited access to stem cell sources restricts their clinical application. To address this issue, we established a simple in vitro epigenetic reprogramming technique in which skin fibroblasts are induced to dedifferentiate into multipotent cells. In this study, human fibroblasts were isolated from circumcised adult foreskin and were reprogrammed by co-culture for 72 h with fish oocyte extract (FOE) in serum-free medium. The cells were then observed and analyzed by immunofluorescence staining, flow cytometry and in vitro differentiation assays. Then FOE-treated human fibroblasts were transplanted by tail vein injection into irradiated mdx mice, an animal model of DMD. Two months after injection, the therapeutic effects of FOE-treated fibroblasts on mdx skeletal muscle were evaluated by serum creatine kinase (CK) activity measurements and by immunostaining and RT-PCR of human dystrophin expression. The results indicated that the reprogrammed fibroblasts expressed higher levels of the pluripotent antigen markers SSEA-4, Nanog and Oct-4, and were able to differentiate in vitro into adipogenic cells, osteoblastic cells, and myotube-like cells. Tail vein injection of FOE-treated fibroblasts into irradiated mdx mice slightly reduced serum CK activity and the percentage of centrally nucleated myofibers two months after cell transplantation. Furthermore, we confirmed human dystrophin protein and mRNA expression in mdx mouse skeletal muscle. These data demonstrated that FOE-treated fibroblasts were multipotent and could integrate into mdx mouse myofibers through the vasculature.
基金:
National Natural Science Foundation of ChinaNational Natural Science Foundation of China [31172170, 81101158]; Foundation for Science & Technology of Yunnan, China [2011HB050, 2013DA004]
第一作者机构:[1]Stem Cell Engineering Laboratory of Yunnan Province, Kunming General Hospital of Chengdu Military Command, Kunming, 650032, P. R. China
通讯作者:
推荐引用方式(GB/T 7714):
Pang Rongqing,Zhu Xiangqing,Geng Jia,et al.IN VITRO AND IN VIVO ANALYSIS OF HUMAN FIBROBLAST REPROGRAMMING AND MULTIPOTENCY[J].CELLULAR & MOLECULAR BIOLOGY LETTERS.2015,20(3):404-417.doi:10.1515/cmble-2015-0024.
APA:
Pang, Rongqing,Zhu, Xiangqing,Geng, Jia,Zhang, Yongyun,Wang, Qiang...&Pan, Xinghua.(2015).IN VITRO AND IN VIVO ANALYSIS OF HUMAN FIBROBLAST REPROGRAMMING AND MULTIPOTENCY.CELLULAR & MOLECULAR BIOLOGY LETTERS,20,(3)
MLA:
Pang, Rongqing,et al."IN VITRO AND IN VIVO ANALYSIS OF HUMAN FIBROBLAST REPROGRAMMING AND MULTIPOTENCY".CELLULAR & MOLECULAR BIOLOGY LETTERS 20..3(2015):404-417