title: Expression level of pluripotent genes in incomplete reprogramming.
abstract: OBJECTIVE
  To compare the expression levels of pluripotent genes among incomplete reprogrammed colonies and induced pluripotent stem cells (iPSCs), to explore the relationship between the expression of pluripotent genes and incomplete reprogramming.
  
  
  METHODS
  Four genes (Oct4, Sox2, Klf4, C-Myc) were introduced into human foreskin fibroblasts (HFFs) by retroviruses. The HFFs were induced to reprogramming. Different forms of colonies were picked up, analyzed, and compared with iPSCs from different aspects, including the morphology of clones, alkaline phosphatase (AP) staining, immuno-fluorescence, and Q-PCR.
  
  
  RESULTS
  In the reprogramming process, different colonies were emerged, some of them exhibited typical human embryonic stem cell morphology (eg., compact colonies, high nucleus-to-cytoplasm ratios, and prominent nucleoli). However, these colonies couldn't maintain these characters after passage. There was an intermediate state, named partially reprogramming. Through analysis and identification, AP staining results were weakly positive, compared with iPSC colonies. The immuno-fluorescence staining demonstrated these colonies just expressed pluripotent protein Oct4. Q-PCR indicated that the expression of exogenous transcription factors was inappropriate, either at a high level or at a low level. Most of the endogenous pluripotency genes were expressed at a low level.
  
  
  CONCLUSIONS
  It may be one of the causes of incomplete reprogramming that the exogenous pluripotent gene is low-expressed or over-expressed, and successful reprogramming may depend on a specific stoichiometric balance of Oct4, Sox2, Klf4 and c-Myc.
authors.affiliation: First Affiliated Hospital of Zhengzhou University, Zhengzhou 450052, Henan Province, China; Affiliated Hospital of Hainan Medical University, Haikou 570102, Hainan Province, China.
authors.forename: Xing
authors.initials: X
authors.lastname: Zhao
authors.affiliation: Affiliated Hospital of Hainan Medical University, Haikou 570102, Hainan Province, China.
authors.forename: Qi
authors.initials: Q
authors.lastname: Li
authors.affiliation: Affiliated Hospital of Hainan Medical University, Haikou 570102, Hainan Province, China.
authors.forename: Wei-Min
authors.initials: WM
authors.lastname: Jiang
authors.affiliation: Affiliated Hospital of Hainan Medical University, Haikou 570102, Hainan Province, China.
authors.forename: Hong-Yan
authors.initials: HY
authors.lastname: Liu
authors.affiliation: Affiliated Hospital of Hainan Medical University, Haikou 570102, Hainan Province, China.
authors.forename: Ning
authors.initials: N
authors.lastname: Ma
authors.affiliation: Affiliated Hospital of Hainan Medical University, Haikou 570102, Hainan Province, China.
authors.forename: Zhi
authors.initials: Z
authors.lastname: Zhou
authors.affiliation: Affiliated Hospital of Hainan Medical University, Haikou 570102, Hainan Province, China.
authors.forename: Lin-Jiang
authors.initials: LJ
authors.lastname: Li
authors.affiliation: Affiliated Hospital of Hainan Medical University, Haikou 570102, Hainan Province, China. Electronic address: Huangyh4180@sohu.com.
authors.forename: Yuan-Hua
authors.initials: YH
authors.lastname: Huang
authors.affiliation: Affiliated Hospital of Hainan Medical University, Haikou 570102, Hainan Province, China. Electronic address: mayanlinma@hotmail.com.
authors.forename: Yan-Lin
authors.initials: YL
authors.lastname: Ma
mesh_terms: D000818:Animals; D002460:Cell Line; D002478:Cells, Cultured; D065150:Cellular Reprogramming / Q000235:genetics*; D002648:Child; D005347:Fibroblasts; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000166:cytology* / Q000502:physiology*; D000090062:Kruppel-Like Factor 4; D008297:Male; D051379:Mice; D008813:Mice, Inbred ICR; D050814:Octamer Transcription Factor-3 / Q000235:genetics / Q000378:metabolism; D012190:Retroviridae / Q000235:genetics; D055746:Stage-Specific Embryonic Antigens / Q000235:genetics / Q000378:metabolism; D014162:Transfection / Q000379:methods
chemical_list: C000715467:KLF4 protein, human; C000715468:Klf4 protein, mouse; D000090062:Kruppel-Like Factor 4; D050814:Octamer Transcription Factor-3; C494173:POU5F1 protein, human; D055746:Stage-Specific Embryonic Antigens; C066584:stage-specific embryonic antigen-4
keywords: Human foreskin fibroblast; Incomplete reprogramming; Induced pluripotent stem cell; Reprogramming