Abstract
The aim of our study was to optimize GCs isolation, transport and a cultivation protocol, which in turn enables monitoring of morphological changes and viability during GCs long-term cultivation in vitro. Human GCs were collected from 28 hormonally stimulated women undergoing in vitro fertilization.
Patients were stimulated by 10.000 IU Pregnyl 36 hrs prior oocytes collection. The cumulus oophorus, obtained transvaginally and oocytes were collected and follicular fluid (FF) contaning GCs was transported to laboratory.
Cells were cultivated in medium supplemented with growth factors (EGF, bFGF), follicle-stimulating hormone (FSH) and 2% fetal calf serum. The significant increase in proliferation of GCs was observed after a single addition of FF to cultivation media, allowing to obtain a large amount of cells with high proliferation potential.