TGF-β_3诱导大鼠前软骨干细胞向软骨细胞特性方向分化及其在KLD-12自组装肽纳米凝胶中的培养  

Induction of Precartilaginous Stem Cells into Chondrogenic Differentiation by TGF-β3 and Cultivation in Self-assembling Peptide KLD-12 Hydrogel

作　　者：孙凯[1] 游洪波[1] 陈安民[1] 郭风劲[1] 祁军[1] 徐志刚[1] 丁然[1] 

机构地区：[1]华中科技大学同济医学院附属同济医院骨外科,武汉430030

出　　处：《华中科技大学学报（医学版）》2010年第1期1-5,共5页Acta Medicinae Universitatis Scientiae et Technologiae Huazhong

基　　金：国家自然科学基金资助项目(No.30571873)

摘　　要：目的探讨软骨组织工程方法中修复软骨缺损理想的种子细胞和支架材料。方法免疫磁珠分离纯化新生大鼠前软骨干细胞(PSCs),分别采用KLD-12自组装肽纳米凝胶三维培养(实验组)和普通培养瓶平面培养(对照组),用转化生长因子β3(TGFβ-3)诱导两组PSCs向软骨细胞特性方向分化。利用RT-PCR、免疫组化等方法测定其向软骨细胞特性方向分化过程中特异性细胞外基质Ⅱ型胶原(collagenⅡ)和聚集蛋白聚糖(aggrecan)表达的情况。结果RT-PCR和免疫组化检测结果表明KLD-12自组装肽纳米凝胶组细胞在诱导7、14 d后均有collagenⅡ和aggrecan表达,且RT-PCR检测结果表明KLD-12自组装肽凝胶组细胞的collagenⅡ和aggrecan mRNA表达水平较对照组高,其差异有统计学意义(均P<0.05)。结论TGFβ-3诱导后的PSCs可向成软骨方向分化,诱导后的PSCs与KLD-12自组装肽凝胶复合有望构建组织工程化软骨,KLD-12自组装肽纳米凝胶是较好的软骨组织工程细胞支架。Objective To evaluate the effects of implanted cell-scaffold composites in the repair of the cartilage defects. Methods Precartilaginous stem cells（PSCs）were isolated and purified by immunomagnetic beads. PSCs were seeded into self-assembling peptide KLD-12 hydrogel and culture flask,then induced into a chondrogenic pathway by TGF-β3. The expression of cartilage specific extracellular matrix Collagen Ⅱ and Aggrecan during chondrogenesis was detected by RT-PCR,and immunohistochemistry. Results Collagen Ⅱ and Aggrecan staining was positive at the 7th and 14th day in KLI〉12 hydrogel culture. RT-PCR revealed that the expression of collagen Ⅱ and Aggrecan mRNA in KLD-12-PSCs was significantly higher than in the PSCs that seeded in culture flask（P〈0.05）. Conclusion PSCs can be induced into chondrogenic differentiation by TC-F-β3. The self-assembling peptide KLD-12 hydrogel loaded with PSCs may be well used for cartilage tissue engineering,and self-assembling peptide KLD-12 hydrogel could be better cell-seeded scaffolds for repair of articular cartilage defects.

关 键 词：前软骨干细胞 转化生长因子Β3 细胞外基质 自组装肽纳米凝胶 

分 类 号：R322.7[医药卫生—人体解剖和组织胚胎学]