Comparison of the culture methods between human cumulus cells and mural granulosa cells / 中国病理生理杂志

ABSTRACT

AIM:To establish an effective method for purification and culture of human cumulus cells(CCs) in vitro,and to compare the characteristics between CCs and mural granulosa cells(MGCs).METHODS:Follicular fluid and cumulus complex from the patients undergoing intracytoplasmic sperm injection were collected.CCs were mechanically cut from cumulus complex and then directly inoculated on a Petri dish, and MGCs were obtained from follicular fluid through density gradient centrifugation.The expression of follicle stimulating hormone receptor(FSHR)was determined by immunofluorescence.The cell growth curves were measured by CCK-8 assay.The secretion of estrogen was detected by ELISA.RESULTS:After incubated for 24 h, the adherence of CCs was observed.CCs and MGCs had similar growth characteristics and FSHR expression.The similar cell growth curves were observed by CCK-8 assay and the results of ELISA showed that they had comparable secretion of estrogen.CONCLUSION:Direct culture of CCs mechanically cutting from cumulus complex is an effective method.CCs had similar growth characteristics,growth curves and secretion of estro-gen to MGCs cultured in vitro and could be a substitutive source of granulosa cell subsets.