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Development of Primordial and Prenatal Follicles from Undifferentiated Somatic Cells and Oocytes in the Hamster Prenatal Ovary In Vitro: Effect of Insulin¹

^a Leland J. and Dorothy H. Olson Center for Women's Health, Division of Reproductive Endocrinology and Infertility, Department of Ob/Gyn and Department of Physiology and Biophysics, University of Nebraska Medical Center, Omaha, Nebraska 68198-4515

Fetal hamster ovaries were cultured for up to 16 days in the presence or absence of various dosages of insulin to evaluate the induction of folliculogenesis in vitro. In the absence of insulin, a few primordial follicle-like structures appeared by the 4th day, and distinct primary follicles (stage 1) appeared by the 12th day of culture. The organelles in the oocytes and adjacent granulosa cells developed along with follicular growth. Moreover, gap junctions between the oocyte and somatic cell plasma membrane also developed as early as 8 days in culture.

In the presence of 0.2 µg/ml insulin, primary follicles developed after 8 days, and ~4% secondary follicles with 2–3 layers of granulosa cells appeared after 16 days of culture. However, higher dosages (> 0.2 µg/ml) of insulin retarded primary follicle formation and induced the formation of primordial follicles with larger oocytes. An increased number of larger oocytes with a few granulosa cells accumulated at the periphery of the ovary. The results indicate that although primordial and primary follicles can develop after 12 days in vitro in the absence of exogenous insulin, the latter is required for timely progression of follicular development through primary and secondary stages.

¹ This work was supported by a grant HD28165 from the National Institute of Child Health and Human Development, NIH, and Olson Foundation for Women's Health, NE.

² Correspondence: Shyamal K. Roy, Departments of OB/GYN and Physiology and Biophysics, University of Nebraska Medical Center, 984515 Nebraska Medical Center, Omaha, NE 68198-4515. FAX: 402 559 6164; skroy{at}mail.unmc.edu

³ Current address: Department of OB/GYN, Division of Reproductive Biology, Stanford University School of Medicine, 300 Pasteur Drive, Room A344, Stanford, CA 94305-5317.

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