Optimization of Culture Conditions for Human Ovarian Stem Cells Using Gelatin-Coated Surfaces

The presence of stem cells in the ovary has been a topic of discussion due to their questionable existence. Isolation of stem cells has been possible by enzymatic digestion; however, the percentage of cells harvested and expanded has not been satisfactory. This could be due to the lack of optimal adhesion provided by the standard commercial culture dishes, which affects the initial attachment and further growth of cells.

Ovarian tissue was collected during pelvic surgery after obtaining written informed consent from four patients. The tissue was exposed to enzymatic digestion using collagenase and hyaluronidase. The resulting cell suspension was cultured in a carbon dioxide incubator in petri dishes, which were either coated with gelatin or uncoated, and monitored serially for growth. The initial cells were counted and further passaged when confluent. The cell number and quality, along with embryoid body formation, were compared between the coated and uncoated dishes.

The initial attachment of cells and further expansion were higher in the dishes that were coated with gelatin compared to the standard non-coated culture dishes.

The addition of gelatin to the culture dishes seems to be beneficial in the culture of stem cells isolated from the human ovarian tissue and their further expansion.