title: Differentiation of human foreskin fibroblast-derived induced pluripotent stem cells into hepatocyte-like cells.
abstract: The aim of this study was to investigate the differentiation potential of induced pluripotent stem cells (iPSCs) derived from human foreskin fibroblasts (HFFs) into hepatocyte-like cells (HLCs). The iPSCs were firstly induced by transduction of OCT4, SOX2, KLF4, and c-MYC into HFFs using retrovirus. Afterwards, expressions of pluripotency factors were identified by semiquantitative reverse transcription-polymerase chain reaction and immunofluorescence staining, and karyotype, embryoid, and teratoma were observed by microscope. Then, iPSCs were gradually differentiated into endoderm cells, hepatic progenitor cells, and mature HLCs by special culture medium. During this process, differentiation efficiency into each kind of cells was evaluated by detecting SOX17, HNF4a, and ALB using flow cytometry, respectively. Besides, enzyme-linked immunosorbent assay was conducted to detect the secretion of ALB in iPSC-induced HLCs and quantitative reverse transcription-polymerase chain reaction was performed to detect the expression levels of hepatocyte-specific genes. The iPSCs were successfully induced by HFFs, which exhibited typical embryonic stem cells morphology, positive alkaline phosphatase staining, normal diploid karyotype, and positive expression of various pluripotency factors. Meanwhile, spherical embryoid and teratoma with 3 germ layers were formed by iPSCs. The iPSCs were consecutively induced into endoderm cells, hepatic progenitor cells and mature HLCs, and the differentiation efficiency was 55.7 ± 2.9%, 45.7 ± 4.8%, and 35.0 ± 3.9%, respectively. Besides, the secretion of ALB and expression of various hepatocyte-specific genes was highly detected in iPSC-induced HLCs. The iPSCs were successfully derived from HFFs and then differentiated into HLCs, which proved a new source for hepatocyte transplantation.
  
  
  CONCLUSIONS
  HFFs were successfully induced into iPSCs by transduction of OCT4, SOX2, KLF4, and c-MYC. Positive expressions of various pluripotency factors were exhibited in HFFs-induced iPSCs. The iPSCs were consecutively induced into endoderm cells, hepatic progenitor cells, and mature HLCs. Various hepatocyte-specific genes were highly expressed in iPSC-induced HLCs.
authors: International Centre for Diagnosis and Treatment of Liver Diseases, 302 Military Hospital of China, Beijing, China.
authors: Jianjun
authors: J
authors: Wang
authors: International Centre for Diagnosis and Treatment of Liver Diseases, 302 Military Hospital of China, Beijing, China.
authors: Ping
authors: P
authors: Zhao
authors: Liver Failure Treatment and Research Center, 302 Military Hospital of China, Beijing, China.
authors: Zhihong
authors: Z
authors: Wan
authors: International Centre for Diagnosis and Treatment of Liver Diseases, 302 Military Hospital of China, Beijing, China.
authors: Xueyuan
authors: X
authors: Jin
authors: International Centre for Diagnosis and Treatment of Liver Diseases, 302 Military Hospital of China, Beijing, China.
authors: Yongqian
authors: Y
authors: Cheng
authors: International Centre for Diagnosis and Treatment of Liver Diseases, 302 Military Hospital of China, Beijing, China.
authors: Tao
authors: T
authors: Yan
authors: International Centre for Diagnosis and Treatment of Liver Diseases, 302 Military Hospital of China, Beijing, China.
authors: Song
authors: S
authors: Qing
authors: International Centre for Diagnosis and Treatment of Liver Diseases, 302 Military Hospital of China, Beijing, China.
authors: Ning
authors: N
authors: Ding
authors: Liver Failure Treatment and Research Center, 302 Military Hospital of China, Beijing, China. xsj302@yahoo.com.cn.
authors: Shaojie
authors: S
authors: Xin
mesh_terms: D000418:Albumins / Q000378:metabolism; D002454:Cell Differentiation; D058732:Embryoid Bodies / Q000166:cytology; D005347:Fibroblasts / Q000166:cytology* / Q000378:metabolism; D005455:Fluorescent Antibody Technique; D052816:Foreskin / Q000166:cytology*; D005786:Gene Expression Regulation; D022781:Hepatocytes / Q000166:cytology* / Q000378:metabolism; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000166:cytology* / Q000378:metabolism; D007621:Karyotyping; D000090062:Kruppel-Like Factor 4; D008297:Male
chemical_list: D000418:Albumins; C000715467:KLF4 protein, human; D000090062:Kruppel-Like Factor 4
keywords: differentiation; hepatocyte transplantation; hepatocyte-like cells; human foreskin fibroblasts; induced pluripotent stem cells