ABSTRACT
Background: One of the well-known causes of subfertility is polycystic ovary syndrome (PCOS). Genetic components play a critical role in the etiology of PCOS. The recognition of differentially expressed genes in PCOS patients might provide a better understanding of the pathophysiology of this syndrome and paves the way for novel therapeutics. Gene expression profiles in cumulus cells (CCs) could be used as biological criteria for embryo competence and their analysis might lead to important molecular information about embryo quality. CALM1, PSMD6, and AK124742 are three well-known genes associated with embryo development. Therefore, the objective of this study was to compare the expression of CALM1, PSMD6, and AK124742 genes in the CCs of infertile PCOS patients with their expression in the CCs of the donor fertile group. Materials and Methods: CCs were collected from the follicular fluid of 33 patients with PCOS as the experimental group and 33 cumulus donor women who were referred to the infertility center for egg donation as the control group. CCs were frozen until genetic testing. The expression of CALM1, PSMD6, and AK124742 genes was detected by real-time polymerase chain reaction. Results: CALM1 and AK124742 gene expressions significantly increased (CALM1 P = 0.003) (AK124742 P = 0.000) and PSMD6 expression significantly decreased (P = 0.002) in the PCOS group compared to the cumulus donor (control) group. Conclusion: Therefore, our research findings suggest that the potential impact of Polycystic Ovary Syndrome (PCOS) on fertility could be attributed to modifications in the expression levels of genes that affect the reproductive.
ABSTRACT
Azoospermia is one of the challenging disorders affecting couples who are afflicted with infertility. Human testisderived cells (hTCs) are suitable candidates for the initiation of in-vitro spermatogenesis for these types of patients. The current study aimed to assess the proliferation of hTCs through the cell culture on the three-dimensional (3D) porous scaffolds. Cells harvested from the testicular sperm extraction (TESE) samples of the azoospermic patients were cultured on the 3D porous scaffolds containing human serum albumin (HSA)/tri calcium phosphate nanoparticles (TCP NPs) for two weeks. The proliferation/viability of the cells was assessed using the MTT assay, along with H and E histological staining method. The MTT assay showed that hTCs could stay alive on this scaffold with 50 and 66.66% viability after 7 and 14 days, respectively. Such viability was not significantly different when compared with cells grown on monolayer flask culture (P>0.05). Therefore, 3D HSA/TCP NPs scaffolds could be used for the reconstitution of the artificial human somatic testicular niche for future applications in regenerative medicine for male infertility.
ABSTRACT
OBJECTIVE: Recent achievements in stem cell biotechnology, nanotechnology and tissue engineering have led to development of novel approaches in regenerative medicine. Azoospermia is one of the challenging disorders of the reproductive system. Several efforts were made for isolation and culture of testis-derived stem cells to treat male infertility. However, tissue engineering is the best approach to mimic the three dimensional microenvironment of the testis in vitro. We investigated whether human testis-derived cells (hTCs) obtained by testicular sperm extraction (TESE) can be cultured on a homemade scaffold composed of electrospun nanofibers of homogeneous poly (vinyl alcohol)/human serum albumin/gelatin (PVA/HSA/gelatin). MATERIALS AND METHODS: In this experimental lab study, human TCs underwent two steps of enzymatic cell isolation and five culture passages. Nanofibrous scaffolds were characterized by scanning electron microscopy (SEM) and Fouriertransform infrared spectroscopy (FTIR). Attachment of cells onto the scaffold was shown by hematoxylin and eosin (H and E) staining and SEM. Cell viability study using MTT [3-(4, 5-dimethyl-2-thiazolyl) -2, 5-diphenyl -2H- tetrazolium bromide] assay was performed on days 7 and 14. RESULTS: Visualization by H and E staining and SEM indicated that hTCs were seeded on the scaffold. MTT test showed that the PVA/HSA/gelatin scaffold is not toxic for hTCs. CONCLUSION: It seems that this PVA/HSA/gelatin scaffold is supportive for growth of hTCs.
ABSTRACT
Human male germ-line stem cells (hmGSCs) and human testis-derived embryonic stem cell-like (htESC-like) cells are claimed to be in vitro pluripotent counterparts of spermatogonial stem cells (SSCs), but the origin and pluripotency of human testis-derived cell cultures are still under debate. The aim of this study was to generate putative pluripotent stem cells in vitro from human testicular sperm-extracted (TESE) samples of infertile men, and to assess their pluripotency and capacity to differentiate. TESE samples were minced, enzymatically disaggregated and dispersed into single-cell or cluster suspensions, and then cultured. Initially, cell clusters resembled those described for hmGSCs and htESC-like cells, and were positive for markers such as OCT4/POU5F1, NANOG, and TRA-2-54. Prolonged propagation of cell clusters expressing pluripotency markers did not thrive; instead, the cells that emerged possessed characteristics of mesenchymal stromal cells (MSCs) such as STRO-1, CD105/EGLN1, CD13/ANPEP, SOX9, vimentin, and fibronectin. KIT, SOX2, and CD44 were not expressed by these MSCs. The multipotential differentiation capacity of these cells was confirmed using Oil Red-O and Alizarin Red staining after induction with specific culture conditions. It is therefore concluded that pluripotent stem cells could not be derived using the conditions previously reported to be successful for TESE samples.