Histone demethylases JHDM1D, PHF2 and PHF8 expression pattern in granulosa cells obtained from patients undergoing IVF procedure during short-term IVC

Abstract

Granulosa cells play an important role in follicle development, maturation, and atresia. They are a cellular source of the two most important ovarian steroids, namely, estradiol and progesterone and are also crucial for bidirectional communication with the oocyte, thus being involved in the regulation of its growth, development and function. Growing body of evidence suggests that granulosa cells cultured in vitro display stemness and transdifferentiation potential. Together with the fact that they can be easily collected during IVF procedures, these properties of GCs may be of particular interest for both regenerative medicine and transplantology. Establishment of in vitro cell culture and its thorough characterization, including molecular, is crucial for future potential utilization of human granulosa cells in design of engineered tissue grafts or cell-based therapies, in particular targeted at female infertility. Nevertheless, the transcriptomic alterations which may occur during in vitro culture of granulosa cells are still largely uncharacterized. The aim of this study was to examine expression changes of three genes encoding histone demethylases which serve as transcription coactivators in short term in vitro cell culture of human granulosa cells. The study groups consisted of 14 patients, aged 18–40 years undergoing in vitro fertilization (IVF). Expression level assessment was performed after 24 h, 48 h, 72 h, 96 h, 120 h, 144 h and 168 h of in vitro primary cell culture utilizing RT-qPCR technique. Upregulation of PHF2 expression in all time points of the culture was observed, whereas the tendency of JHDM1D and PHF8 was mainly to decrease in expression level. Further study on a larger population would be required in order to confirm the presented tendencies. Running title: Expression pattern of selected histone demethylases in human granulosa cells

Expression Profile of New Gene Markers and Signaling Pathways Involved in Immunological Processes in Human Cumulus-Oophorus Cells

“…Total RNA purity was estimated by using the 260/280 nm absorption ratio. Samples from 12 patients with an 260/280 nm absorbance ratio greater than 1.8 were used for RT-qPCR analysis [11,12]. For microarray expression, the 12 trials obtained were randomly divided into 2 equal groups which contained RNA from 8 patients.…”