慢病毒对大鼠骨髓间充质干细胞生物学特性的影响  

Effects of lentiviral vector on biological characteristics of rat bone marrow mesenehymal stem cells

作　　者：王萍[1] 田玉科[1] 田学愎[1] 程秋菊[1] 陈莎莎 宋珂[2] 罗婷[1] 

机构地区：[1]华中科技大学同济医学院附属同济医院麻醉学研究室,武汉市430030 [2]华中科技大学同济医学院附属同济医院口腔科,武汉市430030

出　　处：《中华麻醉学杂志》2009年第5期415-418,共4页Chinese Journal of Anesthesiology

基　　金：国家自然科学基金资助项目（30471660）;湖北省卫生厅科研基金项目（JX2807）

摘　　要：目的探讨慢病毒对大鼠骨髓间充质干细胞（rMSCs）生物学特性的影响。方法将含有绿色荧光蛋白基因的慢病毒感染第3代rMSCs。观察感染的rMSCs的形态学；感染的rMSCs与FITC标记的抗大鼠CD29抗体、PE标记的抗大鼠CD90抗体及APC标记的抗大鼠CD45抗体孵育后，立即检测细胞表面CD29、CD90、CD45的表达水平；感染的rMSCs在成骨诱导培养基中培养21d时，观察骨细胞的形成情况；感染的rMSCs在成脂肪诱导培养基中培养14d时，观察脂肪细胞的形成情况；感染的rMSCs培养1、2、3、4、5、6、7、8d时测定细胞活力；感染的rMSCs在琼脂培养基中培养21d时，观察细胞克隆的形成情况；将感染的rMSCs接种于BALB／c—nu／nu裸鼠背部，观察接种后42d内接种部位肿瘤的形成情况。结果感染的rMSCs的形态学无改变，表达CD29^＋CD90^＋CD45^-，具有形成骨细胞和脂肪细胞的能力，细胞活力无明显改变，无细胞克隆形成，接种裸鼠后接种部位未见肿瘤形成。结论慢病毒对大鼠骨髓间充质干细胞生物学特性无影响。Objective To investigate the effects of lentiviral vector（LV）on the biological characteristics of the rat bone marrow mesenchymal stem cells （rMSCs）. Methods rMSCs derived from the third passage were infected by the LV containing a green fluorescent protein gene. The morphology of the LV-infected rMSCs was observed. The expression of CD29, CD90 and CD45 was detected immediately after the LV-infected rMSCs were incubated with FITC labeled anti-rat CD29 antibody, PE labeled anti-rat CD90 antibody and APC labeled anti-rat CD45 antibody. The LV-infected rMSCs were cultured in osteogenic induction media and osteocyte formation was observed at 21 d of culture. The LV-infected rMSCs were cultured in adipogenic induction media and adipocyte formation was observed at 14 d of culture. The viability of the LV-infected rMSCs was detected at 1, 2, 3, 4, 5, 6, 7 and 8 d of culture. The LV-infected rMSCs were cultured in soft agar culture media and cell clones were observed at 21 d of culture. The LV-infected rMSCs were inoculated into the back of BALB/e-nu/nu nude mice and the tumorigenesis was observed within 42 d after inoculation. Results The morphology of the LV-infected rMSCs had no change. The LV-infected rMSCs were positive for CD29, CD90 and negative for CD45 and differentiated into osteoblasts and adipocytes. The cell viability did not change. There was no soft agar colony formation. No tumor was formed at the inoculation site. Conclusion Lentiviral vector dose not affect the biological characteristics of the rMSCs.

关 键 词：慢病毒属 间质干细胞移植 生物学特性 疼痛 

分 类 号：R3[医药卫生—基础医学]