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Kit ligand and c-Kit are expressed during early human ovarian follicular development and their interaction is required for the survival of follicles in long-term culture

¹ Karolinska Institutet, Department of Clinical Science, Intervention and Technology. Karolinska University Hospital, Stockholm, Sweden, ² Program for Developmental and Reproductive Biology, Biomedicum Helsinki and Department of Bacteriology and Immunology, Haartman Institute, University of Helsinki, Helsinki, Finland, ³ Jorvi Hospital, Helsinki University Central Hospital, Espoo, Finland, ⁴ Infertility Clinic, The Family Federation of Finland, Helsinki, Finland and ⁵ Department of Reproductive Science and Medicine, Division of Paediatrics, Obstetrics and Gynaecology, Imperial College School of Medicine, Hammersmith Hospital, London, UK

Correspondence should be addressed to O Hovatta, Karolinska Institutet, Department of Clinical Science, Intervention and Technology, Karolinska University Hospital Huddinge, SE 14186 Stockholm, Sweden; Email: Inger.Britt.Carlsson{at}klinvet.ki.se

The receptor tyrosine c-Kit and its cognate ligand, c-Kit ligand (KL, stem cell factor, SCF), are involved in ovarian follicular development in several animal species. We studied the expression of KL and c-Kit using in situ hybridization and immunohistochemistry in donated human ovarian cortical tissue. The KL transcripts were expressed in granulosa cells of primary follicles, whereas the expression of c-Kit was confined to the oocyte and granulosa cells in primary and secondary follicles. We employed an ovarian organ culture using firstly serum-containing and then serum-free medium to study the effects of KL and an anti-c-Kit antibody, ACK2, on the development and survival of ovarian follicles in vitro. Culture of ovarian cortical slices for 7 days resulted in a 37% increase in the number of primary follicles and a 6% increase in secondary follicles. The proportion of viable follicles decreased in all cultures. The addition of KL (1, 10 and 100 ng/ml) into the culture media did not affect the developmental stages of the follicles or the proportion of atretic follicles. Inclusion of ACK2 (800 ng/ml) in the culture medium significantly increased the proportion of atretic follicles on days 7 (49 vs 28% in control cultures) and 14 (62 vs 38%) of culture. In conclusion, c-Kit and KL are expressed in human ovaries during follicular development. Blocking the c-Kit receptor induces follicular atresia. The KL/c-Kit signaling system is likely to control the survival of human ovarian follicles during early follicular development.

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