【摘要】 [目的]研究体外培养大鼠骨髓间质干细胞的超微结构。[方法]采用常规技术对SD鼠骨髓间质干细胞进行体外培养传代、鉴定、诱导分化和透射电镜分析。[结果]流式细胞仪检测,细胞表达CD29和CD44,不表达CD11b和CD45;经5-氮杂胞苷和两性霉素B诱导分化后结蛋白和肌球蛋白染色阳性。透射电镜观察有巨噬样、内皮样和成纤维样三类基本细胞,巨噬样细胞表面有胞质突起,胞浆含电子密度高的结晶样包涵体;内皮样细胞靠胞膜边有吞饮小泡,胞浆中有电子密度高的内皮特殊小体;成纤维样细胞呈梭形,较幼稚,细胞器简单,有丰富的糖原和核糖体;经诱导分化的肌样细胞胞浆靠胞膜缘可见无细胞器的条状肌丝区带;细胞间存在胞膜局部紧密连接和胞质突起接合。[结论]传代贴壁生长的为骨髓间质干细胞;超微结构分析显示早期幼稚细胞发育的特征,但各有特点;在不同培养条件诱导下,细胞具备向肌肉和脂肪组织分化的潜能。本研究为进一步了解和掌握骨髓间质干细胞的生物学特性和临床应用打下基础。更多还原

【Abstract】 [Objective] To study the ultrastructure of rat marrow-derived mesenchymal stem cells (MSC) cultured in vitro. [Methods] The MSC of SD rat were cutured,passaged,induced and differentiated in vitro using routine culture technique, and evaluated by FACScan flow cytometer and analyzed by Hitachi H-600 transmission electron microscope(TEM). [Results] FACScan showed that the cells expressed antigens of CD29 and CD44 but not CD11b and CD45. The cells showed positive stain with desmin and myoglobin after induction with 5-azacytidine and amphotericin B. Three kinds of cells including macrophage-like,endothelium-like and fibroblast-like cells were observed with TEM. A lot of surface cytoplasm process and plasmal electron-dense crystalloid inclusion were found in the macrophage-like cells. The endothelium-like cells were found with pinocytosis vesicle beside the membrane and electron-dense specific granule in the plasm. The fibroblast-like cells were in the shape of fusiform and immature, and its organell was simple. There were a lot of glycogen and ribosome in the plasm. Stripform zone of myofilament without any organells in the margin of myoid cells membrane were observed after induction and differentiation. Membrane structures of partial tight junction and the joints of cytoplasm process with intercells were found. [Conclusion] The passaged cells are MSC. The study of ultrastructure displays that MSC have the characters of earlier immature cells, but each has its own behavior. Under different conditions, MSC have potency of differentiation into muscle and fatty tissues. The study provides us better background for further understanding of the biocharacteristics and clinical application of MSC.更多还原