大鼠骨髓基质细胞超微结构观察及胶质源性酸性蛋白的表达

Ultrastructure of Rat Bone Marrow Stromal Cell and Its Glial Fibrillary Acidic Protein Expression

目的:体外分离培养大鼠骨髓基质细胞(BMSCs),通过超微结构观察和免疫组织化学检测探讨细胞生物学特性。方法:收集幼年大鼠骨髓细胞,利用贴壁培养法纯化扩增。培养细胞至第3代,相差显微镜及苏木精-伊红染色,扫描电镜和透射电镜观察细胞生长状态和细胞形态;免疫细胞化学染色检测细胞表面神经标记物巢蛋白(Nestin),神经特异性烯醇化酶(NSE),胶质源性酸性蛋白(GFAP)。结果:电镜下细胞表面见大量微绒毛结构,一些细胞有单个或多个突起样结构。多数细胞膜完整,胞质内存在丰富的细胞器;胞核大而明显,染色质分布正常;细胞间连接较少,可见缝隙连接。免疫细胞化学检测GFAP阳性细胞为(15.05±3.92)%。结论:第3代BMSCs具有生长旺盛,增值能力强,分化程度低的特点;部分细胞可表达GFAP。

Aim： To explore the biological characteristics of BMSC by observing its ultrastructure and detecting its neural markers, for further research on differentiation of BMSCs in vitro. Methods. Bone marrow cells（BM）samples were obtained aseptically from healthy juvenile rat femur and cells of the third passage were then selected by plastic adhesion. Phase contrast microscope, HE （hematoxylinand eosin）staining, and electron microscope were applied to observe morphocytological characteristic. Cells were finally determined by immunocytochemical stain with Nestin, neuron special enolase（NSE）, and glial fibrillary acidic protein（GFAP）, and positive cells were statistically counted. Results： Under election microscope, plenty of microvilli were seen on the surface of the third passage of BMSCs. A great of cells had integrant membrane and abundant organelles in the cytoplasma, and the nuclei were large and chromatin were normally distribution. Gap junction was also found between cells. Immunohistochemical stain indicated that GFAP positive cells were （15.05 ± 3.92）% of the total cells, while no Nestin and NSE positive cells were found. Condusion： BMSCs at passage 3 proliferated rapidly and the degree of their differentiation was low. A few of them expressed GFAP naturally.