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

摘要：

目的　探讨碱性成纤维细胞生长因子对大鼠阴茎肌源性干细胞体外增殖的影响。方法　本试验自2020年12月至2021年6月在滕州市中心人民医院精准实验室完成。取健康雄性Wister大鼠5只，体质量（200±50）g，取其阴茎海绵体并采用酶消化法分离肌源性干细胞，应用密度梯度离心法和差速贴壁法进行纯化。取第2代肌源性干细胞分为两组进行培养，实验组：200 µl生长培养基+肌源性干细胞，分别用终浓度为10 µg/L、30 µg/L、50 µg/L、70 µg/L、90 µg/L外源性碱性成纤维细胞生长因子进行干预；对照组：加入200 µl生长培养基+肌源性干细胞。分别于培养24 h、48 h、72 h、96 h、120 h、144 h加入四甲基偶氮唑蓝（MTT）溶液。应用免疫细胞化学法鉴定大鼠阴茎肌源性干细胞，MTT比色法检测细胞增殖情况。统计学方法采用独立样本t检验、Tukey检验。结果　（1）肌源性干细胞多呈Sca-1阳性反应［表达量为（79.90±1.82）%］，少数呈Desmin阳性反应［表达量为（68.60±0.72）%］。（2）两组肌源性干细胞均随培养时间的延长而明显增殖（均P<0.01），出现显著促增殖效应的培养时间均为96 h。（3）与对照组比较，实验组各浓度碱性成纤维细胞生长因子对肌源性干细胞均有明显的促增殖效应；当浓度从10～70 µg/L递增时，促增殖作用也随之均显著增强（均P<0.01），至70 µg/L时其促增殖作用接近顶峰。结论　外源性碱性成纤维细胞生长因子可促进体外培养的大鼠阴茎肌源性干细胞增殖，且呈浓度-时间依赖性增加，96 h与70 µg/L为最佳的体外培养时间和浓度。

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