LiCl诱导大鼠脂肪基质细胞成骨分化研究  

LiCl induced differentiation of adipose-derived stromal cells of rat to osteoblasts

作　　者：高倩[1] 赵丽丽[1] 宋子仪[1] 王少璞[1] 张雅卿[1] 杨公社[1] 

机构地区：[1]西北农林科技大学动物科技学院,陕西杨凌712100

出　　处：《西北农林科技大学学报（自然科学版）》2011年第1期15-20,28,共7页Journal of Northwest A&F University(Natural Science Edition)

基　　金：国家自然科学基金项目(30871787);国家大学生创新性实验计划项目(081071208)

摘　　要：【目的】探讨体外培养条件下,LiCl诱导大鼠脂肪基质细胞向成骨细胞分化的潜能。【方法】采用Ⅰ型胶原酶消化组织块法分离获得大鼠脂肪基质细胞,将其分为处理组和对照组,分别用25 mmol/L LiCl和25mmol/L NaCl处理3周,采用形态学、SQ RT-PCR、免疫荧光染色、碱性磷酸酶染色及茜素红染色等方法,检测大鼠脂肪基质细胞向成骨细胞分化的能力。【结果】大鼠脂肪基质细胞在LiCl诱导24 h后,细胞形态由成纤维样变成方块状;诱导3周后,SQ RT-PCR检测发现,细胞表达碱性磷酸酶(AP)、Ⅰ型胶原(ColⅠ)和骨γ羧基谷氨酸(Bglap)等成骨细胞特异性基因,与处理0 d相比,差异达极显著水平(P<0.01),碱性磷酸酶和茜素红染色均呈阳性,其中茜素红染色阳性率可达60%以上;免疫荧光染色检测结果显示,LiCl处理3周后细胞表达成骨细胞特异性蛋白OSX和OPN,阳性率均达70%左右。对照组碱性磷酸酶和茜素红染色均呈阴性,不表达成骨细胞相关基因及蛋白。【结论】大鼠脂肪基质细胞在25 mmol/L LiCl处理下,可分化为成骨样细胞,且表达成骨细胞标志基因及相关蛋白。【Objective】 The study was to investigate adipose-derived stromal cells（ADSCs） of rat differentiation to osteoblasts induced by LiCl in vitro.【Method】 ADSCs were obtained by isolating digestion of collagenase Ⅰ.The treatment group and the control group were treated with 25 mmol/L LiCl and 25 mmol/L NaCl for 3 weeks respectively.The differentiation ability was investigated by morphology,SQ RT-PCR,innumofluorescence,alkaline phosphatase staining,alizarin red stain and statistical analysis.【Result】 ADSCs changed to square from fibroblast-like phenotype after induction by LiCl for 24 h.After 3 weeks,the expression of alkaline phosphatase,ColⅠ and Bglap were detected by Quantitative RT-PCR,and the expression level increased significantly compared with that of 0 day.The alkaline phosphatase staining and alizarin red stain showed positive,with the positive rate of alizarin red stain more than 60%.And the expression of OSX and OPN were detected by immunofluorescence staining,with positive rate 70% or so.While the control group showed negative for alkaline phosphatase staining and alizarin red stain,and didn＇t express the markers of osteoblasts.【Conclusion】 In a word,ADSCs could differentiate osteoblasts with induction of 25 mmol/L LiCl,and expressed many of osteoblasts-specific markers.

关 键 词：脂肪基质细胞 氯化锂 成骨分化 大鼠 

分 类 号：Q813.11[生物学—生物工程]