豚鼠脂肪间充质干细胞:一种适用于耳科实验研究的理想干细胞

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:观察豚鼠脂肪间充质干细胞成骨、成脂肪、成神经细胞分化的潜能,并探讨其应用于耳科实验的可能。方法:取豚鼠脂肪,分离脂肪间充质细胞培养至第3代,检测第1～10天细胞增殖吸光度值;流式细胞仪检测细胞的免疫表型;并行成骨细胞诱导、成脂肪细胞诱导、成神经细胞诱导及鉴定。结果与结论:脂肪间充质细胞增殖的生长曲线近似"S"形,传代后经过3d的潜伏期,4d进入对数生长期,8d进入平台期;锥虫蓝染色细胞拒染率94%～98%,从P0～P5细胞的总产量扩增了约3600倍;其细胞免疫表型CD29表达率86.3%,CD44表达率55.5%,CD45表达率为0.4%。成骨细胞诱导后经特异性染色可见矿化结节形成;成脂肪细胞诱导后经油红O染色胞浆内可见脂质滴;成神经细胞诱导后细胞形态与神经细胞类似,Nestin、GFAP、NF200染色均呈阳性。提示豚鼠脂肪间充质细胞在体外易于分离、培养及扩增,具有间充质干细胞的"干性",是一种适用于耳科实验研究的理想干细胞。

Guinea pig adipose-derived mesenchymal stem cells:Ideal stem cells for experimental studies of otology

BACKGROUND:Up to date,most of the animal experiments about stem cells research on the otology use the stem cells of other animal species for the guinea pig model,however,the research of guinea pig-derived stem cells is rarely reported.OBJECTIVE:To explore the differentiated potential of the guinea pig adipose mesenchymal stem cells into osteoblasts,adipocyte,nerve cells,and the application possibility for experimental study of otology.METHODS:Guinea pigs adipose tissue was isolated and cultured to the 3rd generation.Absorbance values of cell proliferation from 1 to 10 days were detected and the immunophenotype of the cells were detected by flow cytometry.Differentiation of osteoblast,adipocyte and nerve cells were verified.RESULTS AND CONCLUSIONS:The growth curve of adipose mesenchymal cell proliferation is similar to S-shaped,the first 3 days was the incubation period after passage and the cell populations entered logarithmic growth phase from the 4th day,after the 8th day they entered into the platform period.The rejection rate of trypan blue staining was 94%-98%,the total cells was amplified by about 3 600 times from primary to passage 5,The expression rate of cells of the immune phenotype CD29 was 86.3%,CD44 was 55.5% and CD45 was 0.4%.After differentiation of osteoblast,specific staining demonstrated the formation of mineralized nodules.After differentiation of adipocyte,lipid droplets could be seen in cytoplasm by oil red O staining.After differentiation of nerve cells,cells morphology was similar to neural cells and the results of Nestin,GFAP,NF200 staining were positive.Guinea pig adipose mesenchymal stem cells was easy to isolate,culture and amplification in vitro.They have the "stem" characteristics of mesenchymal stem cells,and they were ideal stem cells for experimental studies of otology.