caa a22 4500 445817429 CHVBK 20180317145233.0 cr unu---uuuuu 170323e20110901xx s 000 0 eng 10.1007/s10815-011-9617-6 doi (NATIONALLICENCE)springer-10.1007/s10815-011-9617-6 Proliferative potential and phenotypic analysis of long-term cultivated human granulosa cells initiated by addition of follicular fluid [Elektronische Daten] [Lenka Bruckova, Tomas Soukup, Benjamin Visek, Jiri Moos, Martina Moosova, Jana Pavelkova, Karel Rezabek, Lenka Kucerova, Stanislav Micuda, Eva Brcakova, Jaroslav Mokry] Purpose: 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 45days with a doubling time of 159 ± 24h. 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. Conclusion: Our present study has demonstrated that GCs can be maintained in vitro for at least 45days and this cell model can be beneficial when studying hormonal regulation associated with follicular maturation and preparation of oocytes for fertilization. Springer Science+Business Media, LLC, 2011 Cultivation nationallicence Follicular fluid nationallicence Granulosa cells nationallicence Phenotype nationallicence bFGF : basic fibroblast growth factor nationallicence COV434 : cell line derived from a human granulosa cell tumor (Sigma-Aldrich) nationallicence DMEM/F12 : dulbecco's modified eagle's medium nationallicence DT : doubling time nationallicence EGF : epidermal growth factor nationallicence FCS : fetal calf serum nationallicence FF : follicular fluid nationallicence FSH : follicle stimulating hormone nationallicence GCs : luteinizing human granulosa cells nationallicence GC8 : one of the isolated GCs from patient with different characters nationallicence IVF : in vitro fertilization nationallicence LH : luteinizing hormone nationallicence PD : the number of population doublings nationallicence Bruckova Lenka Department of Analytical Chemistry, Faculty of Chemical Technology, University of Pardubice, Pardubice, Czech Republic aut Soukup Tomas Department of Histology and Embryology, Charles University in Prague, Medical Faculty in Hradec Kralove, Simkova 870, 500 38, Hradec Kralove, Czech Republic aut Visek Benjamin Department of Histology and Embryology, Charles University in Prague, Medical Faculty in Hradec Kralove, Simkova 870, 500 38, Hradec Kralove, Czech Republic aut Moos Jiri Department of Obstetrics and Gynecology, Charles University in Prague, 1st Medical Faculty and General Teaching Hospital in Prague, Prague, Czech Republic aut Moosova Martina Department of Obstetrics and Gynecology, Charles University in Prague, 1st Medical Faculty and General Teaching Hospital in Prague, Prague, Czech Republic aut Pavelkova Jana Department of Obstetrics and Gynecology, Charles University in Prague, 1st Medical Faculty and General Teaching Hospital in Prague, Prague, Czech Republic aut Rezabek Karel Department of Obstetrics and Gynecology, Charles University in Prague, 1st Medical Faculty and General Teaching Hospital in Prague, Prague, Czech Republic aut Kucerova Lenka Department of Clinical Genetics, Teaching Hospital in Hradec Kralove, Hradec Kralove, Czech Republic aut Micuda Stanislav Department of Pharmacology, Charles University in Prague, Medical Faculty in Hradec Kralove, Prague, Czech Republic aut Brcakova Eva Department of Pharmacology, Charles University in Prague, Medical Faculty in Hradec Kralove, Prague, Czech Republic aut Mokry Jaroslav Department of Histology and Embryology, Charles University in Prague, Medical Faculty in Hradec Kralove, Simkova 870, 500 38, Hradec Kralove, Czech Republic aut Journal of Assisted Reproduction and Genetics Springer US; http://www.springer-ny.com 28/10(2011-09-01), 939-950 1058-0468 28:10<939 2011 28 10815 https://doi.org/10.1007/s10815-011-9617-6 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1007/s10815-011-9617-6 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Bruckova Lenka Department of Analytical Chemistry, Faculty of Chemical Technology, University of Pardubice, Pardubice, Czech Republic aut NATIONALLICENCE 700 1- Soukup Tomas Department of Histology and Embryology, Charles University in Prague, Medical Faculty in Hradec Kralove, Simkova 870, 500 38, Hradec Kralove, Czech Republic aut NATIONALLICENCE 700 1- Visek Benjamin Department of Histology and Embryology, Charles University in Prague, Medical Faculty in Hradec Kralove, Simkova 870, 500 38, Hradec Kralove, Czech Republic aut NATIONALLICENCE 700 1- Moos Jiri Department of Obstetrics and Gynecology, Charles University in Prague, 1st Medical Faculty and General Teaching Hospital in Prague, Prague, Czech Republic aut NATIONALLICENCE 700 1- Moosova Martina Department of Obstetrics and Gynecology, Charles University in Prague, 1st Medical Faculty and General Teaching Hospital in Prague, Prague, Czech Republic aut NATIONALLICENCE 700 1- Pavelkova Jana Department of Obstetrics and Gynecology, Charles University in Prague, 1st Medical Faculty and General Teaching Hospital in Prague, Prague, Czech Republic aut NATIONALLICENCE 700 1- Rezabek Karel Department of Obstetrics and Gynecology, Charles University in Prague, 1st Medical Faculty and General Teaching Hospital in Prague, Prague, Czech Republic aut NATIONALLICENCE 700 1- Kucerova Lenka Department of Clinical Genetics, Teaching Hospital in Hradec Kralove, Hradec Kralove, Czech Republic aut NATIONALLICENCE 700 1- Micuda Stanislav Department of Pharmacology, Charles University in Prague, Medical Faculty in Hradec Kralove, Prague, Czech Republic aut NATIONALLICENCE 700 1- Brcakova Eva Department of Pharmacology, Charles University in Prague, Medical Faculty in Hradec Kralove, Prague, Czech Republic aut NATIONALLICENCE 700 1- Mokry Jaroslav Department of Histology and Embryology, Charles University in Prague, Medical Faculty in Hradec Kralove, Simkova 870, 500 38, Hradec Kralove, Czech Republic aut NATIONALLICENCE 773 0- Journal of Assisted Reproduction and Genetics Springer US; 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