大鼠脂肪干细胞体外培养及成骨诱导分化研究  

Isolation,culture and osteogenic differentiation of rat adipose-derived stem cells

作　　者：梁丽华[1] 李龙[2] 赵建辉[3] 孙道才[1] 刘乃彬[1] 宋应亮[1] 焦铁军[1] 

机构地区：[1]第四军医大学口腔医学院种植科,西安710032 [2]第四军医大学口腔医学院口腔颌面外科,西安710032 [3]第四军医大学西京医院全军整形外科研究所

出　　处：《实用口腔医学杂志》2012年第3期289-293,共5页Journal of Practical Stomatology

基　　金：国家自然科学基金资助项目(编号:81170984)

摘　　要：目的:观察SD大鼠脂肪干细胞(adipose-derived stem cells,ASCs)体外培养的基本生物学特性、鉴定其诱导成骨分化潜能。方法:无菌条件下取6周龄SD大鼠腹股沟处脂肪组织,0.1%I型胶原酶消化,分离培养。显微镜下观察细胞形态;采用MTT比色法绘制细胞生长曲线;用流式细胞仪检测细胞表面标志物;取第3代细胞行成骨和成脂诱导培养,分别用碱性磷酸酶(ALP)、茜素红染色和油红O染色鉴定成骨诱导效果。结果:从SD大鼠脂肪组织中分离获得的ASCs呈长梭形或多角形,呈集落生长;细胞生长曲线呈"S"型,具有较强的增殖能力;流式细胞仪分析示:CD29、CD90高表达,CD34、CD45低表达;成骨诱导后ALP染色及茜素红染色阳性,成脂诱导后油红O染色阳性。结论:分离培养的细胞为ASCs,具有成骨潜能。Objective： To study the biological characteristics and osteogenic potency of adipose-derived stem cells（ASCs）.Methods： Cells were isolated from the bilateral inguinal fat pads by collagenase I digestion and cultured.Cell morphology was observed under the phase contrast microscope.The cell proliferation was tested by MTT assay.The surface markers of the cells were analyzed by flow cytometry.Passage 3 cells were cultured in osteogenic and adipogenic inducing medium for 7,28 and 21 days respectively,and verified with ALP staining,Alizarin red staining and Oil Red O staining respectively.Results： The cells presented fibroblast-like or polygon shape.The growth curve was like ＂S＂ shape reflecting strong proliferative ability.ASCs expressed CD29 and CD90,but not CD34 and CD45.The ALP staining,Alizarin red staining and Oil Red O staining showed positive reaction.Conclusion： ASCs can be cultured from SD rat fat tissue and can be induced to osteogenic differentiation.

关 键 词：ASCs 体外培养 成骨诱导分化 

分 类 号：R329[医药卫生—人体解剖和组织胚胎学]