关键词:

肌源性干细胞, 大鼠阴茎, 外源性碱性成纤维细胞生长因子, 增殖, 浓度, 培养时间

Abstract:

Objective　To investigate the effect of basic fibroblast growth factor on the proliferation of rat corpus cavernosal muscle-derived stem cells. Methods　This experiment was completed in the Precision Laboratory of Tengzhou Central People's Hospital from December 2020 to June 2021. The penis was taken from five healthy male Wister rats [(200±50) g] under aseptic conditions, and the muscle-derived stem cells were isolated by enzymatic digestion and purified by density gradient centrifugation and differential adhesion. The second generation of muscle-derived stem cells were selected and were divided into two groups. The experimental group: 200 µl of growth medium + the second generation of muscle-derived stem cells were treated with 10 µg/L, 30 µg/L, 50 µg/L,70 µg/L, and 90 µg/L of exogenous basic fibroblast growth factor; the control group added 200 µl of growth medium + muscle-derived stem cells. The methyl thiazolyl tetrazolium (MTT) solution was added after 24 h, 48 h, 72 h, 96 h, 120 h, and 144 h of culture. The rat corpus cavernosal muscle-derived stem cells were identified by immunocytochemistry, and the cell proliferation was detected by MTT assay. Independent sample t test and Tukey test were used. Results　(1)Most of the muscle-derived stem cells showed Sca-1 positive reaction, with the expression level of (79.90±1.82) %, while a few showed Desmin positive reaction, with the expression level of (68.60±0.72) %. (2)The muscle-derived stem cells in both groups proliferated significantly with the prolongation of culture time (all P<0.01), and the culture time for significant proliferative effect was 96 h. (3)Compared with the control group, various concentrations of basic fibroblast growth factor in the experimental group had obvious proliferative effect on muscle-derived stem cells. When the concentration increased from 10 to 70 µg/L, the proliferative effects were also significantly enhanced (all P<0.01), and the proliferative effect reached the peak when the concentration was at 70 µg/L. Conclusion　Exogenous basic fibroblast growth factor can promote the proliferation of rat corpus cavernosal muscle-derived stem cells in vitro in a concentration-time dependent manner, with 96 h and 70 µg/L as the optimal in vitro culture time and concentration.

Key words:

Muscle-derived stem cells, Rat corpus cavernosal, Exogenous basic fibroblast growth factor, Proliferation, Concentration, Culture time