摘要
目的分离培养扩增大鼠脂肪源间充质干细胞(ADSCs),以活体标记并鉴定其分化潜能,了解ADSCs的X连锁凋亡抑制蛋白(XIAP)基因修饰的可行性。方法无菌条件下取大鼠一侧腹股沟脂肪组织,Ⅰ型胶原酶消化法分离培养ADSCs,胰酶消化法传代扩增。检测细胞分化为脂肪细胞、软骨细胞及成骨细胞的潜能,转染XIAP表达质粒进入ADSCs,通过Western blotting等方法检测XIAP的表达能力。结果 ADSCs呈长梭形漩涡样生长,细胞流式鉴定显示CD29、CD44、CD90、CD105均呈高表达,并在特定诱导剂下分化为脂肪细胞、软骨细胞或成骨细胞。XIAP转染后显像经XIAP基因修饰的脂肪间充质干细胞在PVDF膜的相应分子质量区域出现相应的条带。结论脂肪源干细胞易于培养和传代扩增,并可活体标记,具有多向分化潜能,可作为组织工程的种子细胞。
Objective To investigate the feasibility of genetically modified X-linked inhibitor of apoptosis protein （XIAP） of rat adipose-derived mesenchymal stem ceils （ADSCs） by isolating and cultivating rat ADSCs in vitro. Methods ADSCs were isolated from rat groin fat pads by collagenase ][ digestion under sterile condition. ADSCs were passaged and amplified with 10% FBS DMEM. The nmlti-differentiation potential of ADSCs was verified by cultivated with differentiation medium. XIAP expression plasmid was transfeeted into ADSCs. The anti-apoptotic ability of XIAP transduetion was detect- ed by Western blotting assay. Results ADSCs were mainly spindle-shaped and whirlpool-shaped arranged. Results of flow eytometry showed that there were higher expressions of CD29, CD44, CD90 and CD105 in ADSCs, which differentiated into lipoeytes, ehondrocytes and osteoblasts under specific conditions. There is XIAP gene modified adipose-derived mesenehy- real stem cells Band in the corresponding molecular mass of PVDF membrane area. Conclusion ADSCs were isolated from rat subcutaneous fat pads and were easily cultivated, passaged and amplified. ADSCs can differentiate into osteoblasts, chon- drocytes and adipoeytes under specific conditions, which are better resource for being used in cell therapy and tissue engi- neering.
出处
《天津医药》
CAS
北大核心
2013年第8期799-801,I0003,共4页
Tianjin Medical Journal
基金
天津市卫生局科技基金重点项目(项目编号:10KG120)
