豚鼠脂肪间充质干细胞向类神经元样细胞的分化

背景:目前关于耳科干细胞研究的动物实验大多是将其他动物种属的干细胞用于豚鼠模型,而对于豚鼠本身来源的干细胞研究报道极少.目的:探寻豚鼠脂肪间充质干细胞向神经细胞元样分化的潜能.方法:取豚鼠脂肪,分离脂肪间充质干细胞培养至第3代,cck-8试剂盒检测第1~10天细胞增殖A值,流式细胞仪检测细胞的免疫表型;成神经诱导并行巢蛋白、胶质纤维酸性蛋白、神经丝蛋白200免疫荧光染色.结果与结论:豚鼠脂肪间充质干细胞增殖的生长曲线近似"S"形,传代后经过3 d的潜伏期,3 d后进入对数生长期,8 d后进入平台期,其细胞的免疫表型CD29表达率86.3%,CD44表达率55.5%,CD45表达率为0.4%,成神经细胞诱导后细胞形态与神经细胞类似,巢蛋白、胶质纤维酸性蛋白、神经丝蛋白200染色均呈阳性.说明豚鼠脂肪间充质干细胞在体外易于分离、培养及扩增,具有向神经细胞元样分化的潜能.

Guinea pig adipose mesenchymal stem cells differentiate into neuron-like cells

BACKGROUND: At present, most of the animal experiments about ear stem cells research are use the stem cells of the other animal species for the guinea pig model. The stem cells from guinea pig was rarely reported. OBJECTIVE: To explore the differentiated potential of the guinea pig adipose mesenchymal stem cells into neuron-like cells. METHODS: Guinea pigs adipose tissue were obtained, the adipose mesenchymal cells were isolated and cultured to the 3rd generation. (1) The absorbance values of cell proliferation from 1 to 10 days were tested by cck-8 kit. (2) The immunophenotype were detected by flow cytometry. (3) Differentiation of neuron-like cells was verified by nestin, glial fibrillary acidic protein (GFAP) and neurofilament protein 200 (NF200) immunofluorescence staining. RESULTS AND CONCLUSION: (1) The growth curve of adipose mesenchymal stem cells proliferation was similar to S-shaped, the first 3 days was the incubation period after passage, the cell populations entered logarithmic growth phase from the 4th day, after the 8th day they entered into the platform period. (2) The expression rate cells immune phenotype CD29 was 86.3%, CD44 was 55.5%, CD45 was 0.4%. (3) After differentiation of nerve cells, the morphology of cells was similar to neural cells, the results of nestin, GFAP and NF200 staining were positive. Guinea pig adipose mesenchymal stem cells are easy to isolation, culture and amplification in vitro. They have the potential to differentiate into neuron-like cells.