‘Heart development and morphogenesis’ is a novel pathway for human ovarian granulosa cell differentiation during long‑term in�vitro cultivation‑a microarray approach

Abstract

Granulosa cells (GCs) have many functions in the endocrine system. Most notably, they produce progesterone following ovulation. However, it has recently been proven that GCs can change their properties when subjected to long-term culture. In the present study, GCs were collected from hyper-stimulated ovarian follicles during in vitro fertilization procedures. They were grown in vitro , in a long-term manner. RNA was collected following 1, 7, 15 and 30 days of culture. Expression microarrays were used for analysis, which allowed to identify groups of genes characteristic for particular cellular processes. In addition, reverse transcription-quantitative polymerase chain reaction (RT-qPCR) was performed to validate the obtained results. Two ontological groups characteristic for processes associated with the development and morphogenesis of the heart were identified during the analyses: ‘Heart development’ and ‘heart morphogenesis’. The results of the microarrays revealed that the highest change in expression was demonstrated by the lysyl Oxidase, oxytocin receptor, nexilin F-actin binding protein , and cysteine-rich protein 3 genes. The lowest change was exhibited by odd-skipped related transcription factor 1, plakophilin 2, transcription growth factor-β receptor 1 , and kinesin family member 3A . The direction of changes was confirmed by RT-qPCR results. In the present study, it was suggested that GCs may have the potential to differentiate towards other cell types under long-term in vitro culture conditions. Thus, genes belonging to the presented ontological groups can be considered as novel markers of proliferation and differentiation of GCs towards the heart muscle cells.

Expression of genes involved in neurogenesis, and neuronal precursor cell proliferation and development: Novel pathways of human ovarian granulosa cell differentiation and transdifferentiation capability in�vitro

“…it has also been suggested that this gene is involved in the formation of the heart and cardiac muscle cells (58). Therefore, its expression has also been discussed in a previous study regarding the differentiation of Gcs to myocardial cells (39). Many scientists believe that neurogenesis also occurs in adult mammals (59)(60)(61)(62)(63)(64).…”

“…The results indicated that these cells have the potential to differentiate towards neurons, as they express neural-differentiation specific genes, providing further proof for their stem-like potential (14,20). a number of studies have reported that it is possible to obtain neuronal-like cells during the differentiation/transdifferentiation of other cell types (5,39,43,52). notably, neural-like cells can be obtained through adipocyte differentiation.…”

“…Previous work. Part of the materials and methods is based on other publications from the same research team, presenting results from the cycle of studies related to human ovarian Gcs (37)(38)(39)(40)(41) of gonadotropins and GnrH antagonist were controlled and recorded for every patient. The follicular fluid (FF) containing oocytes and GCs was collected and transferred to a qualified embryologist who extracted all oocytes from the fluid and passed it on to the further stages of iVF.…”

“…once adherent cells were >90% confluent, they were detached with 0.05% trypsin-EDTA (Gibco; Thermo Fisher Scientific, Inc.) for 1-3 min and counted using a fluorescence automatic cell counter (adaM-Mc; nanoenTek america, inc.). Gcs were then cultivated for 30 days; total rna was isolated after 1, 7, 15 and 30 days (38,39,43). The medium was replaced every 72 h. cell morphology was observed after 1, 7, 15 and 30 days of culture under an inverted light microscope (olympus iXc73; olympus corporation).…”

Ovarian follicular cells - living in the shadow of stemness cellular competence

“…Research groups are going into individual avenues on how GCs, VSELs and OSCs could potentially be used in clinical applications. One of the main interests in using stem cells found in the ovaries is for restoring female fertility, but regeneration of tissues like the heart are also being researched and stem cells may also be used as a model for greater understanding of cell differentiation and epigenetics [15,16].…”