摘要
目的探讨大鼠骨髓基质细胞(mesenchymal stem cells,MSCs)的体外培养生物学特性及其定向诱导分化能力。方法从大鼠双侧股骨中分离获取MSCs,体外培养扩增纯化后传代于塑料培养皿中,相差显微镜观察其形态学特点,MTT法测定绘制传代骨髓间充质干细胞生长曲线,经成骨诱导液,20%新生牛血清(fetal bovineserum,FBS),10mmol/L β-甘油磷酸钠,8～10mmol/L地塞米松,50mg/L维生素C,神经细胞诱导液,0.06mg/L神经生长因子(nerve growth factor,NGF),20%FBS。对传至3～5代的细胞进行体外诱导分化,采用免疫细胞化学法对诱导后的细胞进行鉴定。结果大鼠MSCs在体外培养中具有增殖、分化潜能,可分化为软骨细胞、神经细胞等。结论 MSCs可在体外大量扩增,具有较强的自我更新及多向分化能力,在适宜的诱导分化条件下,可诱导为神经干细胞。
Objective To discuss the condition of culture and differentiation of rat bone marrow mesenchymal stem cells （MSCs）in vitro. Methods The MSCs were isolated primarily from femurs of Sprague- Dawley adult rat under anesthesia, which were planted into plastic petridishes and cultured. Morphological characters of MSCs were examined by phase contrast microscopy, and the growth curve of cultured MSCs was drawn via MTT results. MSCs were treated with osteogenetic inductor （20% fetal bovine serum, 10mmol/L 13 - sodium glycerophosphate, 8 - 10mmol/L dexamethasone, 50mg/L vitamin C） ;Neural inductor,O. 06mg/L nerve growth factor（NGF）, 20% fetal bovine serum . The 3rd to 5th passage of MSCs were induced in vitro. Immunocytochemistry was used to identify cell types. Results BMSC could be expanded and generated when they were cultured in vitro. These cells grew rapidly and differentiated into neuron- like ceils and osteoblast. Conclusion Bone marrow -derived MSCs can be isolated and cultured in vitro , which have the better potentiality of proliferation and multi - directional differentiation. MSCs can differentiate into neuron if the culture condition is appropriate.
出处
《河北医科大学学报》
CAS
2011年第3期252-255,共4页
Journal of Hebei Medical University
基金
河北省保定市科技局科学技术研究与发展计划项目(06F04-11)
河北省卫生厅科研基金项目(20090175)
关键词
骨髓细胞
细胞培养技术
大鼠
bone marrow cells
cell cuhure techniques
rats
