title: [Establishment of feeder-free culture system of human parthenogenetic embryonic stem cells].
abstract: OBJECTIVE
  To establish a safe, effective, and economic feeder'-free culture system which is suitable for the culture of human parthenogenetic embryonic stem cells (hPESCs) in vitro.
  
  
  METHODS
  hPESCs were cultured with mTeSR 1 medium (control group) and human foreskin fibroblasts-conditional medium (hFFs-CM) (experimental group). The growth status of hPESCs in both feeder-free culture systems were observed with inverted microscope. Alkaline phosphatase (ALP) analysis and karyotype analysis were used to study the biological characteristics of hPESCs. The expression of hPESCs pluripotent marker Oct-4 was analyzed by RT-PCR. Differentiation experiment in vivo and in vitro was applied to observe the differentiation potential of hPESCs into three germ layers.
  
  
  RESULTS
  hPESCs had regular morphology with difficulty in differentiation in both culture systems. No obvious difference was observed in morphology and expansion speed of hPESCs between 2 groups. After subcultured for 15 passages in vitro, hPESCs in 2 groups could maintain normal female diploid karyotype 46, XX and pluripotency. The expression of Oct-4 mRNA was positive in 2 groups. hPESCs in 2 groups could form embryonic body in differentiation experiment in vitro and could develop into teratomas containing three germ layers in nude mice.
  
  
  CONCLUSIONS
  Feeder-free culture system of hFFs-CM can sustain the growth of hPESCs and keep hPESCs undifferentiated state for long. A feeder-free culture system of hPESCs is successfully established, which can support the growth of hPESCs, reduce the contamination from animals, decrease the cost of culture, and satisfy the clinical large-scale application.
authors.affiliation: Department of Pathology, 'Genetic Diagnosis Center, the First People's Hospital of Yunnan Province, Kunming Yunnan, 650032, PR China.
authors.forename: Rui
authors.initials: R
authors.lastname: Liang
authors.forename: Zhiqiang
authors.initials: Z
authors.lastname: Wang
authors.forename: Tianxing
authors.initials: T
authors.lastname: Chen
authors.forename: Jing
authors.initials: J
authors.lastname: Zhu
authors.forename: Shu
authors.initials: S
authors.lastname: Zhu
authors.forename: Ying
authors.initials: Y
authors.lastname: Li
authors.forename: Long
authors.initials: L
authors.lastname: Yang
authors.forename: Baosheng
authors.initials: B
authors.lastname: Zhu
mesh_terms: D000818:Animals; D015415:Biomarkers / Q000378:metabolism; D018929:Cell Culture Techniques / Q000379:methods*; D002454:Cell Differentiation; D018920:Coculture Techniques; D017077:Culture Media, Conditioned; D053595:Embryonic Stem Cells / Q000166:cytology* / Q000378:metabolism; D061252:Feeder Cells; D005260:Female; D005347:Fibroblasts / Q000166:cytology* / Q000378:metabolism; D052816:Foreskin / Q000166:cytology*; D006801:Humans; D007621:Karyotyping; D008297:Male; D051379:Mice; D008819:Mice, Nude; D050814:Octamer Transcription Factor-3 / Q000378:metabolism; D010312:Parthenogenesis; D039904:Pluripotent Stem Cells / Q000166:cytology / Q000378:metabolism
chemical_list: D015415:Biomarkers; D017077:Culture Media, Conditioned; D050814:Octamer Transcription Factor-3; C494173:POU5F1 protein, human