Lipoprotein signaling & cholesterol metabolism associated gene expression in human ovarian granulosa cells during IVF
Sawarkar, Sarthak Sanjay
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Background: In vitro fertilization (IVF) is an inefficient process. Better biomarkers are required to improve the selection of oocytes and embryos and thereby increase the efficiency of this procedure. Follicular cholesterol levels have been shown to be associated with embryo morphology parameters and may be useful predictors of IVF outcomes. This study examines gene expression in human ovarian granulosa cells (GCs) comparing GCs from follicles which generate highly-fragmented embryos with GCs from follicles which generate healthy embryos. Methods and Materials: Human ovarian granulosa cells were harvested from patients during standard clinical IVF protocols. Total RNA was extracted from these GCs and converted into complementary DNA (cDNA). cDNA was then utilized to run focused RT-PCR arrays to examine the expression 84 lipoprotein signaling and cholesterol metabolism associated genes. The fold changes in gene expression were determined between GC samples based on associated embryo morphology parameters, particularly embryo fragmentation. Results: Two single follicles were aspirated from a 180 patients which, following IVF protocols, generated 130 embryos. Of these 130 embryos only 21 of the associated follicles GCs provided sufficient total RNA for focused array analysis as highly variable amounts of total RNA (3 μg–17μg) were extracted. Focused PCR array analysis indicated significant upregulation in 14 Lipoprotein signaling and Cholesterol metabolism genes in GC samples with poorest, most highly fragmented embryos. No significantly down-regulated genes were observed. Discussion: Multiple cholesterol metabolism associated genes are upregulated in ovarian granulosa cells derived from follicles which ultimately generate poor quality embryos. Further study of these genes and their protein products could provide clues to understanding the mechanism responsible for the association between follicular cholesterol metabolism and female fertility.