Isolation and preliminary characterization of pig primordial follicles
- 1 November 1989
- journal article
- Published by Bioscientifica in Reproduction
- Vol. 87 (2) , 561-571
- https://doi.org/10.1530/jrf.0.0870561
Abstract
An enzymic method for recovering primordial follicles from the pig ovary consists of incubating cortical slices for 2 h with 0.025% collagenase 1A. An average of 185,000 or 419,000 primordial follicles per ovary were recovered from ovaries collected in Cambridge and Kansas, respectively. Following a discontinuous Percoll gradient, primordial follicles can be separated from contaminating somatic cells by mouth pipette or a micromanipulator to collect 100-1500 follicles but for large scale recovery of approximately 30,000 follicles flow cytometry is recommended. Two types of primordial follicles can be distinguished by electron microscopy: peripheral clusters of small oocytes with an incomplete investment of pregranulosa cells and a deeper region of individual oocytes surrounded by a complete layer of pregranulosa cells. The viability of the purified primordial follicles is attested by their ability to synthesize proteins for at least 12 h after incubation with [35S]methionine. Moreover, the primordial follicles showed several polypeptide bands in common with mature oocytes especially with Mr of 60,000-90,000 but with considerable differences from somatic cells.Keywords
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