title: Proliferative potential and phenotypic analysis of long-term cultivated human granulosa cells initiated by addition of follicular fluid.
abstract: OBJECTIVE
  The aim of this study was to develop and optimize a strategy for long-term cultivation of luteinizing human granulosa cells (GCs).
  
  
  METHODS
  GCs were cultivated in DMEM/F12 medium supplemented with 2% fetal calf serum. In vitro proliferation of GCs was supported by follicular fluid as well as FSH and growth factors.
  
  
  RESULTS
  The cultured GCs were maintained for 45 days with a doubling time of 159 ± 24 h. GCs initiated by the addition of follicular fluid and cultivated under low serum conditions reached 10 ± 0.7 population doublings. GCs maintain the typical phenotypic expression and the telomere length according to specific culture conditions.
  
  
  CONCLUSIONS
  Our present study has demonstrated that GCs can be maintained in vitro for at least 45 days and this cell model can be beneficial when studying hormonal regulation associated with follicular maturation and preparation of oocytes for fertilization.
authors.affiliation: Department of Analytical Chemistry, Faculty of Chemical Technology, University of Pardubice, Pardubice, Czech Republic.
authors.forename: Lenka
authors.initials: L
authors.lastname: Bruckova
authors.forename: Tomas
authors.initials: T
authors.lastname: Soukup
authors.forename: Benjamin
authors.initials: B
authors.lastname: Visek
authors.forename: Jiri
authors.initials: J
authors.lastname: Moos
authors.forename: Martina
authors.initials: M
authors.lastname: Moosova
authors.forename: Jana
authors.initials: J
authors.lastname: Pavelkova
authors.forename: Karel
authors.initials: K
authors.lastname: Rezabek
authors.forename: Lenka
authors.initials: L
authors.lastname: Kucerova
authors.forename: Stanislav
authors.initials: S
authors.lastname: Micuda
authors.forename: Eva
authors.initials: E
authors.lastname: Brcakova
authors.forename: Jaroslav
authors.initials: J
authors.lastname: Mokry
mesh_terms: D018929:Cell Culture Techniques; D049109:Cell Proliferation; D002478:Cells, Cultured; D005260:Female; D005434:Flow Cytometry; D015571:Follicular Fluid; D006107:Granulosa Cells / Q000166:cytology*; D006801:Humans; D007621:Karyotyping; D010641:Phenotype; D016615:Telomere; D013997:Time Factors