title: Serum replacement with a growth factor-free synthetic substance in culture medium contributes to effective establishment of mouse embryonic stem cells of various origins.
abstract: OBJECTIVE
  To evaluate whether serum replacement with growth factor-free synthetic substances contributed to the effective establishment of embryonic stem (ES) cells.
  
  
  METHODS
  Randomized, prospective model study.
  
  
  METHODS
  Gamete and stem cell biotechnology laboratory at Seoul National University in Korea.
  
  
  METHODS
  F1 (C57BL6 x DBA2) mice.
  
  
  METHODS
  Blastocysts of different origins were cultured in serum-replaced media.
  
  
  METHODS
  Embryonic stem cell establishment.
  
  
  RESULTS
  Eight batches of ES cells were established from colony-forming inner cell mass cells after the replacement of fetal bovine serum (FBS) with synthetic knockout serum replacement (KSR) in mkDMEM. The established cells were positive for ES cell markers and formed both embryoid bodies in vitro and teratomas in vivo, but the established cell batches and control (transformed) ES cells responded differently to the culture media. Higher levels of cell viability were detected after the replacement with the 75:25 FBS-KSR mixture than with any other mixtures, and a gradual decrease in viability was detected as the KSR volume ratio was increased. The 75:25 FBS-KSR mixture-containing medium supported ES cell establishment of outbred ICR, F1, and F2 of C57BL6/DBA2; F1 parthenogenetic and ES cell-complemented tetraploid blastocysts; and single ES-cell cultures.
  
  
  CONCLUSIONS
  A serum-replaced medium could be used for effective ES-cell establishment of various origins.
authors.affiliation: Department of Food and Animal Biotechnology, Seoul National University, Seoul, Korea.
authors.forename: Seung Tae
authors.initials: ST
authors.lastname: Lee
authors.forename: Se Woong
authors.initials: SW
authors.lastname: Oh
authors.forename: Dae Yong
authors.initials: DY
authors.lastname: Kim
authors.forename: Jae Yong
authors.initials: JY
authors.lastname: Han
authors.forename: Shin Yong
authors.initials: SY
authors.lastname: Moon
authors.forename: Jeong Mook
authors.initials: JM
authors.lastname: Lim
mesh_terms: D000818:Animals; D021461:Animals, Outbred Strains; D015415:Biomarkers / Q000378:metabolism; D001755:Blastocyst / Q000166:cytology / Q000187:drug effects; D002417:Cattle / Q000097:blood; D002470:Cell Survival; D002478:Cells, Cultured; D003470:Culture Media / Q000138:chemical synthesis* / Q000737:chemistry / Q000494:pharmacology*; D004622:Embryo, Mammalian / Q000166:cytology*; D005260:Female; D005312:Fetal Blood; D006133:Growth Substances / Q000032:analysis; D008297:Male; D051379:Mice; D008815:Mice, Inbred Strains; D013234:Stem Cells / Q000166:cytology* / Q000187:drug effects* / Q000378:metabolism / Q000502:physiology
chemical_list: D015415:Biomarkers; D003470:Culture Media; D006133:Growth Substances