不同浓度血清对大鼠骨骺干细胞向软骨细胞分化的诱导效果比较  

Effect of different-concentration serum on the differentiation of precartilaginous stem cells into chondrocytes in rats

作　　者：张树威[1] 陈安民[1] 宋登新[1] 谢伯臻[1] 王江[1] 祁军[1] 易磊[1] 郭风劲[1] 

机构地区：[1]华中科技大学同济医学院附属同济医院骨科,湖北省武汉市430030

出　　处：《中国组织工程研究与临床康复》2008年第34期6617-6620,共4页Journal of Clinical Rehabilitative Tissue Engineering Research

基　　金：国家自然科学基金(30571872);教育部博士点基金资助项目[教技发中心函(2005)216]~~

摘　　要：背景:血清成分复杂,其中不仅存在促细胞生长因子,同时也存在细胞生长抑制因子,因此合适的血清浓度对于体外培养细胞的增殖和分化至关重要。目的:比较不同浓度的胎牛血清对大鼠骨骺干细胞向软骨细胞分化的影响。设计、时间及地点:细胞学体外观察,于2007-03/10在华中科技大学同济医学院附属同济医院矫形外科研究室完成。材料:纯系清洁级新生24h内的SD大鼠8只,用于制备骨骺干细胞。胎牛血清为Gibco公司产品。方法:免疫磁珠分离纯化FGFR-3阳性的骨骺干细胞,稳定传至第3代后,分别用含1.25%,2.5%,5%,10%胎牛血清的软骨诱导培养基进行干预,诱导14d。主要观察指标:BrdU-ELISA法检测细胞增殖活性。应用免疫荧光染色和RT-PCR法检测细胞Ⅱ型胶原的表达。结果:骨骺干细胞向软骨细胞诱导14d后,10%胎牛血清组细胞增殖活性显著高于1.25%,2.5%,5%胎牛血清组(P<0.05);10%胎牛血清组Ⅱ型胶原免疫荧光染色阳性程度及Ⅱ型胶原mRNA的表达均明显强于其他3组。结论:含10%胎牛血清的软骨诱导液更有利于骨骺干细胞向软骨细胞方向分化。BACKGROUND： Serum has complicated component, including cell growth factor and cell growth inhibitory factor. Suitable concentration of serum is important for proliferation and differentiation of in vitro cultured cells. OBJECTIVE： To compare the effects of different concentrations of fetal bovine serum on the differentiation of precartilaginous stem cells （PCSCs） into chondrocytes in rats. DESIGN, TIME AND SETTING： The cytology in vitro experiment was performed at the Research Room of Orthopaedic Surgery, Tongji Hospital, Tongji Medical College, Huazhong University of Science ＆ Technology from March to October 2007. MATERIALS： Eight 24-hour old Sprague Dawley （SD） rats were used for harvesting PCSCs. Fetal bovine serum was purchased from Gibco. METHODS： Immunomagnetic separation was used to segregate rat PCSCs labeled with fibroblast growth factor receptor-3. At the third passage, PCSCs were induced to differentiate into chondrocytes by chondrogenic culture media containing 1.25%, 2.5%, 5% or 10% of fetal bovine serum （FBS） for 2 weeks. MAIN OUTCOME MEASURES： BrdU-ELISA assay was used to detect the cell proliferation activity in all groups. Immunofluorescence staining and reverse transcription-polymerase chain reaction （RT-PCR） were adopted to test expression of type Ⅱ collagen. RESULTS： At 14 days of induction, cells of 10% FBS group had higher proliferation activity than 1.25%, 2.5%, 5% FBS groups （P 〈 0.05）. Immunofluorescence staining and RT-PCR showed the expression of type Ⅱ collagen and type Ⅱ collagen mRNA expression were significantly stronger in the 10% FBS group compared with the 1.25%, 2.5%, 5% FBS groups. CONCLUSION： The chondrogenic culture media, which contained 10% FBS, is more useful in promoting the proliferation and differentiation of PCSCs into chondrocytes in vitro.

关 键 词：骨骺干细胞 血清浓度 分化 软骨细胞 

分 类 号：R394.2[医药卫生—医学遗传学]