Date

4-2020

Project Type

URC Presentation

Department

Molecular, Cellular and Biomedical Sciences

College or School

COLSA

Class Year

Senior

Major

Biochemistry, Molecular and Cellular Biology

Faculty Research Advisor

Paul Tsang

Abstract

The adult mammalian ovary contains follicles at various stages of development, from primordial to antral. Follicles of all sizes house oocytes and somatic cells that support oocyte maturation through the production of regulatory factors. We have shown that somatic granulosa cells from small and large follicles produce CCN1, which is involved in the formation of new blood vessels. In the present study, our goal was to determine the effect of serum on CCN1 expression in granulosa cells from small follicles. To do so, bovine granulosa cells were obtained from follicles (<5mm) and seeded into T-25 flasks containing DMEM medium supplemented with 1% FBS, 1 ng/mL follicle stimulating hormone (FSH), and 10 ng/mL insulin. At 100% confluency, cells were split into 6-well plates until 80-90% confluency was reached. During this process, 2 of 4 cultures were contaminated. In the 2 remaining cultures, granulosa cells were serum-starved for 2 hours before they were treated with 1%, 5%, or 10% FBS for 2 hours. Then, quantitative polymerase chain reaction was used to determine CCN1 expression. Clearly, granulosa cells from follicles <5mm produced CCN1, and all FBS concentrations appeared to increase CCN1 expression. However, the magnitude of response to FBS differed between the 2 experiments. In a corollary experiment, granulosa cells were not serum-starved, but were maintained with 1% FBS. When they were treated with 10% FBS, CCN1 expression appeared to be higher than in granulosa cells concurrently maintained with 1% FBS. The magnitude of response to FBS appeared to differ between the starved and un-starved cells. Overall, more replicates are needed to confirm these observations.

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