ABSTRACT
Background: Sheep industry has taken steps toward transforming itself into a more efficient and competitive field. There are many varieties of sheep breeds in the world that each of them serves a useful purpose in the economies of different civilizations. Ghezel sheep is one of the Iranian important breeds that are raised for meat, milk and wool. Field of spermatogonial cell technologies provides tools for genetic improvement of sheep herd and multiple opportunities for research. Spermatogonial cells are the only stem cells capable of transmitting genetic information to future generations
Methods: This study was designed to extend the technique of isolation and in vitro proliferation of spermatogonial cells in Ghezel sheep
Results: Isolated cells were characterized further by using specific markers for type A spermatogonia, including PLZF. Also, sertoli cells were characterized by vimentin which is a specific marker for sertoli cells. After 10 days of co-culture, viability rates of the cells was above 94.7%, but after the freezing process the viability rates were 74 percent
Conclusion: In this study, a standard method for isolation and in vitro proliferation of spermatogonial stem cells in Ghezel sheep was developed
ABSTRACT
Background: spermatogonial stem cells [SSCs] are undifferentiated cells which are highly reproducible and expandable. Several studies have been conducted to reproduce these cells in culture. They used growth factors, hormones and different feeder cells to improve survival and proliferation of SSCs
Objective: this study was conducted to evaluate the effects of follicular stimulating hormone [FSH] on gene expression of fibroblast growth factor [FGF2] and glial cell-derived neurotrophic factor [GDNF] in Sertoli cells
Materials and Methods: sertoli cells and SSCs were isolated from 3-5 month-old calves. Bovine testicular cells were cultured for 15 days with or without FSH. Identification of these cells was confirmed by immunocytochemistry analysis. Colony formation of SSCs was evaluated using an inverted microscope. The gene expression of FGF2 and GDNF and the gene markers bcl6b, thy-1, and C-kit were evaluated using the quantitative RT-PCR technique
Results: the results indicated that FSH increased colonization of SSCs. the expression of GDNF, FGF2, and markers of undifferentiated spermatogonia was increased following culture in control and FSH groups [p<0.05], this increase was more in FSH group. Conversely, the expression of C-kit was decreased in both groups [p<0.05]
Conclusion: the results showed that FSH can increase the self-renewal of SSCs in vitro via upregulation of GDNF and FGF2 expression in Sertoli cells
ABSTRACT
Background: Niche cells, regulating Spermatogonial Stem Cells [SSCs] fate are believedto have a reciprocal communication with SSCs. The present study was conductedto evaluate the effect of SSC elimination on the gene expression of Glial cell line-Derived Neurotrophic Factor [GDNF], Fibroblast Growth Factor 2 [FGF2] and KitLigand [KITLG], which are the main growth factors regulating SSCs development andsecreted by niche cells, primarily Sertoli cells
Methods: Following isolation, bovine testicular cells were cultured for 12 days on extracellularmatrix-coated plates. In the germ cell-removed group, the SSCs were removedfrom the in vitro culture using differential plating; however, in the controlgroup, no intervention in the culture was performed. Colony formation of SSCs wasevaluated using an inverted microscope. The gene expression of growth factors andspermatogonia markers were assessed using quantitative real time PCR
Results: SSCs colonies were developed in the control group but they were rarely observedin the germ cell-removed group; moreover, the expression of spermatogoniamarkers was detected in the control group while it was not observed in the germ cellremovedgroup, substantiating the success of SSCs removal. The expression of Gdnfand Fgf2 was greater in the germ cell-removed than control group [p<0.05], whereasthe expression of Kitlg was lower in the germ cell-removed than control group [p<0.05]
Conclusion: In conclusion, the results revealed that niche cells respond to SSCs removalby upregulation of GDNF and FGF2, and downregulation of KITLG in order to stimulateself-renewal and arrest differentiation
ABSTRACT
Spermatogonial Stem Cell [SSC] technologies provide multiple opportunities for research in the field of biotechnology and regenerative medicine. The therapeutic use of Embryonic Stem Cells [ESCs] is restricted due to severe ethical and immunological concerns. Therefore, we need a new pluripotent cell type. Despite well-known role of germ cells in the gametogenesis, some facts apparently show their multipotentiality. In the present study, bovine SSCs were co-cultured with Sertoli cell for 7 days. Sertoli cells and SSCs were identified by Vimentin and Oct-4 immunocytochemical staining method, respectively. In order to differentiate SSCs into osteoblasts, we used consecutive inducer media without separation of the colonies. We characterized osteoblasts using Alizarin red staining