Evaluation of stemness marker expression in bovine ovarian granulosa cells

Abstract

The objective of this study was to assess the stemness marker expressions (Oct4, Nanog, and Sox2) of granulosa cells (GCs) collected from bovine ovarian follicles and in vitro expansion. The single bovine ovarian follicles were isolated and categorized into 4 groups according to their diameter including group A (<2 mm), group B (2-3 mm), group C (3-4 mm), and group D (>4 mm). Quantitative reverse transcriptase polymerase chain reaction (qRT-PCR) and immunostaining were applied to evaluate the stemness marker expression of bovine GCs from ovarian follicles. We also estimated the stemness marker transcript expressions of GCs during in vitro expression by qRT-PCR. qRT-PCR analysis demonstrated that fresh GCs from bovine ovarian follicles expressed the stemness markers (Oct4, Nanog, Sox2). These markers were down-regulated during antral stage follicular development. We also estimated stemness marker transcript expressions of GCs which were isolated and in vitro expanded from ovarian follicles of group A. The qRT-PCR results showed that Oct4 and Sox2 transcript expressions were reduced during in vitro expansion while Nanog transcript was not expressed.

The Stemness of Human Ovarian Granulosa Cells and the Role of Resveratrol in the Differentiation of MSCs—A Review Based on Cellular and Molecular Knowledge

“…Granulosa cells express vimentin [ 18 ], while luteinized GC populations were also reported to express Oct-4 [ 14 , 40 ], further reinforcing suggestions of resemblance to MSC populations. Oct-4 is one of the main regulatory proteins in cell differentiation and self-renewal.…”

“…Along with gradual loss of GC function markers i.e., FSHR, aromatase, throughout the time of their in vitro culture, the expression of markers characteristic for mesenchymal stem cells (MSC) phenotype increases. GCs were successfully differentiated into a variety of cell types, including neurons, osteoblasts and chondrocytes [ 40 ]. GCs also undergo cell division whilst in contact with neighboring cells, without the presence of substratum, a property normally only seen in a stem cell phenotype [ 13 ].…”

Bone morphogenetic protein 15 induces differentiation of mesenchymal stem cells derived from human follicular fluid to oocyte‐like cell

“…Surveys, such as the one conducted by Ding et al (2016), imply a potent stem cell niche into immature ovarian follicles. Previous studies on the multipotency in the follicular antrum cells revealed that some possess characteristics of mesenchymal stem cells (MSCs; Hoang et al, 2019). Ovarian stem cells have opened a new prospect for a better understanding of human oogenesis, infertility preservation, and treatment.…”

Human Granulosa Cells—Stemness Properties, Molecular Cross-Talk and Follicular Angiogenesis

“…Ovarian follicular cells exhibit stem-like potential and present phenotype changes and multipotency during long-term in vitro cultures. Under specific culture conditions eventually leading to successful differentiation into multiple cell types, these stem cells can lose GC functional markers including FSHR and aromatase, and express markers characteristic of mesenchymal stem cell phenotypes [51]. Although the expression of stem cell markers in GCs including OCT-4, Nanog and SOX-2 have been described, their expression varies between different species and maturational stages of GCs [52].…”