大鼠骨髓MSCs体外分离培养及多向分化的实验研究

目的：建立骨髓MSCs体外分离培养体系，并进行多向分化诱导，以证实其多向分化潜能，为MSCs进一步的临床应用研究提供实验依据。方法：用密度梯度离心法分离大鼠骨髓MSCs，并对其形态学特征进行观察。用诱导剂对MSCs向成骨细胞、神经元样细胞和成肌细胞进行诱导分化，并进行形态学观察和免疫细胞化学检测。结果：用密度梯度离心法成功分离获得了高纯度的骨髓MSCs，细胞增殖活性强。经骨向诱导后，ALP和矿化结节染色阳性；免疫细胞化学染色提示神经向诱导神经元特异性标志蛋白NSE和NF200，成肌细胞标志蛋白Deamin和Connexin-43阳性表达。结论：采用密度梯度离心法成功建立了大鼠骨髓MSCs体外分离和培养体系；并能够向成骨细胞、神经元样细胞和成肌细胞分化。

An Experimental Study of Isolation and Pluripotential of Rat Bone Marrow Stem Cells

Objective:To establish an in vitro isolation and culture system of rat bone marrow stem cells and to verify their pluripotential. Methods:Density gradient centrifugation was used to isolate bone marrow MSCs from SD rats and their morphological characteristics were examined under microscope. The MSCs were also undergone osteogenic, neurogenic and myogenic induction and cytochemical and immunocytochemical staining were performed to verify their multipotential. Results:High purity of bone marrow MSCs was successfully obtained and they showed good proliferation ability. Both ALP activity and mineraliztion node staining were positive in MSCs after osteogenic differentiation and MSCs specially expressed neuron markers, NSE and NF200, and myoblast markers, desmin and connexin-43, after neurogenic and myogenic differentiation, respectively. Conclusion:The isolation and culture system of rat bone marrow MSCs was successfully established and the MSCs sustained their osteogenic, neurogenic and myogenic differentiation potential in vitro.