Abstract:

BACKGROUND: To establish a rapid and effective method to obtain sufficient spermatogonial stem cells that can meet the clinical need is urgent to be solved in the spermatogonial stem cell transplantation.
OBJECTIVE: To study the effect of rhodiola polysaccharide on the proliferation of spermatogonial stem cells in vitro.
METHODS: Under sterile conditions, spermatogonial stem cells and Sertoli cells were isolated from the testis of mice, and spermatogonial stem cells were seeded onto the feed layer of Sertoli cells. Then, the co-cultured cells were assigned into experimental group 1 (simple cell culture medium), experimental group 2 (cell culture medium containing 150 mg/L rhodiola polysaccharide) and experimental group 3 (cell culture medium containing 150 mg/L rhodiola polysaccharide, 1 U/L leukemia inhibitory factor and 10 μg/L glial cell line-derived neurotrophic factor). After 7 days of co-culture, flow cytometry was used to detect cell proliferation in vitro, and cell viability and positive expression of GFRa-1, Thy-1 and C-kit were calculated.
RESULTS AND CONCLUSION: After 7 days of co-culture, the cells grew rapidly and presented with colony and thyrsiform growth, and the number of cell masses increased significantly, all of which were in line with the proliferative features of spermatogonial stem cells. The GFRa-1, Thy-1 and C-kit proteins were expressed in the cell membrane and cytoplasm, mainly in the cell membrane. The viability of spermatogonial stem cells and positive expression of GFRa-1 and Thy-1 were ranked as follows: experimental group 3 > experimental group 2 > experimental group 1, and there were significant differences between groups (P < 0.05). The positive expression of C-kit had no difference between experimental groups 1 and 2, but it was significantly higher in the experimental group 3 than the other two groups (P < 0.05). These findings indicate that rhodiola polysaccharide used alone or combined with leukemia inhibitory factor and glial cell line-derived neurotrophic factor can enhance the proliferative ability of spermatogonial stem cells in vitro. 
中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程