Stem Cells
Vahid Akbarinejad; Parviz Tajik; Mansoureh Movahedin; Reza Youssefi
Volume 8, Issue 1 , March 2017, , Pages 7-13
Abstract
Testosterone is believed to play a significant role in spermatogenesis, but its contribution to the process of spermatogenesis is not completely understood. Given that extracellular matrix (ECM) facilitates differentiation of spermatogonial stem cells (SSCs) during culture, the present study was conducted ...
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Testosterone is believed to play a significant role in spermatogenesis, but its contribution to the process of spermatogenesis is not completely understood. Given that extracellular matrix (ECM) facilitates differentiation of spermatogonial stem cells (SSCs) during culture, the present study was conducted to elucidate whether testosterone contribute to the permissive effect of ECM on SSCs differentiation. In experiment 1, testosterone production was measured in testicular cells cultured for 12 days on ECM or plastic (control). In experiment 2, testosterone production was assessed in testicular cells cultured on ECM or plastic (control) and exposed to different concentrations of hCG. In experiment 3, the gene expression of factors involved in testosterone production was analyzed. Testosterone concentration was lower in ECM than in the control group in experiment 1 (p < 0.05). In experiment 2, testosterone concentration was increased in response to hCG in both groups but cells cultured on ECM were more responsive to hCG than those cultured on plastic (p < 0.05). In the experiment 3, qRT-PCR revealed the inhibitory effect of ECM on the gene expression of steroidogenic acute regulatory protein (StAR) (p < 0.05). Nevertheless, the expression of LH receptor was greater in ECM-exposed than in unexposed cells (p < 0.05). In conclusion, the present study showed that inhibiting the expression of StAR, ECM could lower testosterone production by Leydig cells during in vitro culture. In addition, the results indicated that ECM could augment the responsiveness of Leydig cells to hCG through stimulating the expression of LH receptor.
Theriogenology
Reza Youssefi; Parviz Tajik; Mansoureh Movahedin; Vahid Akbarinejad
Volume 7, Issue 4 , December 2016, , Pages 275-280
Abstract
Enrichment of cell suspension with germ cells prior to injection into recipient seminiferous tubules is of importance in spermatogonial stem cells (SSCs) transplantation. Knock-out serum replacement (KSR) has been reported to enhance the proliferation of murine SSCs and human embryonic stem cells. The ...
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Enrichment of cell suspension with germ cells prior to injection into recipient seminiferous tubules is of importance in spermatogonial stem cells (SSCs) transplantation. Knock-out serum replacement (KSR) has been reported to enhance the proliferation of murine SSCs and human embryonic stem cells. The aim of the present study was to investigate the effect of KSR versus fetal bovine serum (FBS) and their interaction on colonization of bovine SSCs in vitro. When FBS (10%) was replaced with KSR (10%), a significant increase in the colonization of SSCs and the expression of Thy1, as marker for enrichment of SSCs, was observed. It was revealed that the lesser proliferative effect of FBS as well as the greater proliferative impact of KSR on SSCs colonization were not irreversible as cells having been cultured with FBS (10%) for three days with low colonization showed high rate of colonization in response to KSR (10%) and cells having been cultured with KSR (10%) with high colonization experienced low rate of colonization in response to FBS (10%). Further, it was shown that FBS did not contain factors inhibiting SSCs colonization and it simply lacked factors essential for SSCs proliferation because the combination of FBS (5%) and KSR (5%) resulted in even greater rate of colonization than did KSR (10%). In conclusion, the present study showed that addition of KSR to culture medium would significantly increase SSCs proliferation.
Stem Cells
Vahid Akbarinejad; Parviz Tajik; Mansoureh Movahedin; Reza Youssefi
Volume 7, Issue 2 , June 2016, , Pages 149-153
Abstract
The receptors 1 and 2 of fibroblast growth factor (FGFR1 and FGFR2, respectively) have been observed in all types of testicular cells. Culture on extracellular matrix (ECM) has been observed to lead to initiation of differentiation in spermatogonial stem cells (SSCs). The present study was carried out ...
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The receptors 1 and 2 of fibroblast growth factor (FGFR1 and FGFR2, respectively) have been observed in all types of testicular cells. Culture on extracellular matrix (ECM) has been observed to lead to initiation of differentiation in spermatogonial stem cells (SSCs). The present study was carried out to investigate whether FGFR1 and FGFR2 play a role in SSCs differentiation. Following isolation, bovine testicular cells were cultured on ECM-coated or uncoated (control) plates for 12 days. The gene expression of THY1, cKIT, FGFR1 and FGFR2 was evaluated using quantitative real-time polymerase chain reaction (PCR). Results related to the gene expression of markers of with undifferentiated (THY1) and differentiated (cKIT) spermatogonia implicated stimulation of self-renewal and differentiation in cells cultured on ECM-coated and uncoated plates, respectively (p < 0.05). Concomitantly, the expression of FGFR2 increased during culture in the ECM group (p < 0.05), whereas it did not change in the control group (p > 0.05). As a result, the gene expression of FGFR2 was greater in the ECM than control group (p < 0.05). Nevertheless, FGFR1 expression did not change during culture in the control and ECM groups (p > 0.05). In conclusion, the present study revealed the potential role of FGFR2 in differentiation of SSCs during culture on ECM.
Reza Narenji Sani; Parviz Tajik; Mohammad Hassan yousefi; Mansoureh Movahedin; Babak Qasemi-Panahi; Shiva Shafiei; Mahmood Ahmadi Hamedani
Volume 4, Issue 1 , March 2013, , Pages 37-41
Abstract
The complex process of spermatogenesis is regulated by various factors. Studies on spermatogonial stem cells (SCCs) have provided very important tool to improve herd genetic and different field. 0.2 to 0.3 percent of total cells of seminiferous tubules is consist of spermatogonial stem cells. To investigate ...
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The complex process of spermatogenesis is regulated by various factors. Studies on spermatogonial stem cells (SCCs) have provided very important tool to improve herd genetic and different field. 0.2 to 0.3 percent of total cells of seminiferous tubules is consist of spermatogonial stem cells. To investigate and biomanipulation of these cells, proliferation and viability rate of cells should be increased in vitro, at first. Follicle stimulating hormone (FSH) has been suggested to play a determinant role in the survival of germ cells in addition to increasing spermatogonial proliferation. In this study, the in vitro effects of FSH on spermatogonial cell colony formation were investigated. Sertoli and spermatogonial cells were isolated from 3-5 months old calves. The identity of the Sertoli cells and spermatogonial stem cells were confirmed through immunocytochemistry and colony morphology, respectively. Co-cultured Sertoli and spermatogonial cells were treated with FSH in different dose of 10, 20 and 40 IU mL-1 FSH, before colony assay. Results indicated that, FSH increased in vitro colonization of spermatogonial cells in comparison with control group. In conclusion, using FSH provided proper bovine spermatogonial stem cell culture medium for in vitro study of these cells.
