(285) Green Eggs and Ham – Development of a Novel System for Primordial Follicle Evaluation in the Porcine Ovary

C. E. Grote1, I. Shofner 1, A. Keller 1, K. Timme 1, A. F. Keating 1* 

1. Department of Animal Science, Iowa State University, Ames, Iowa 

* Corresponding author; akeating@iastate.edu 


Abstract Text: Primordial follicle behavior in the ovary is fundamentally linked to maintaining proper ovarian health and reproductive function. Assessing biological impacts of local growth factors, toxicant exposures, disease, and the normal hormonal milieu on primordial follicles remains constrained due to logistical constraints of primordial follicle isolation. Thus, primordial follicle analysis has relied largely on histological and morphological assessments, coupled with follicle counting as a proxy of impacts on their function. The goal of this study was to develop a method to isolate primordial follicles from ovarian tissue and evaluate biomarkers therein using image-based flow cytometry. Using both mice and porcine ovaries, ovarian stromal tissue was digested using collagenase and primordial follicles were isolated based on selection via an antibody-based isolation strategy. In addition, size exclusion gates of less than 10 µm and more than 50 µm were employed to collect only those sized between. Successful primordial follicle isolation was achieved. Image-based flow cytometry identified that the isolated follicles were positively stained for the protein that served as the isolation target.  This study demonstrates that this is a useful tool for studying biological processes in primordial follicles that may be applicable across species. This work was funded by the Iowa Pork Producers Association and Iowa State University Incentive funding.