摘要
目的 :通过重建端粒酶活性延长胎儿肌肉源间充质干细胞寿命 ,并对其成神经潜能进行研究 ,为组织工程神经修复提供种子细胞。方法 :将人端粒酶催化亚基 (hTERT)基因通过脂质体转染法导入胎儿肌肉源间充质干细胞 ,RT PCR检测hTERTmRNA的表达 ,TRAP PCR检测细胞端粒酶活性。用bFGF诱导已重建端粒酶活性的肌肉源间充质干细胞向神经细胞分化 ,免疫荧光及免疫印迹法检测分化情况。结果 :转染hTERT的胎儿肌肉源间充质干细胞能稳定表达端粒酶活性。转染后传 75代的细胞经bFGF诱导仍维持着自我更新及向神经细胞分化的潜能 ,且无恶性转化倾向。结论 :重建端粒酶活性可延长胎儿肌肉源间充质干细胞寿命并维持自我更新及成神经潜能 。
Objective:To extend the proliferative life span,assess the differentiation potential of telomerase immortalized mesenchymal stem cells (MSCs) derived from human fetal muscle cells and provide `seed cells' for tissue engineering.Methods: The pGRN145 plasmids encoding human telomerase reverse transcriptase (hTERT) were introduced into the human fetal muscle derived MSCs (MMSCs) by cationic lipid reagent mediated transfection in vitro. Telomerase activity was analyzed by TRAP PCR assay.The hTERT positive MMSCs were then induced to differentiate into neural cells,and the results were assayed by immunofluorescence and Western blot.Results:Stable telomerase activity could be detected in the transfected MMSCs and still had the capacity for self renewal and the ability to differentiate into neuronal cells when expanded for more than 70 cell doublings without tendency of malignant transformation.Conclusion:The life span of MMSCs could be prolonged and the capacity for self renewal and the neural differentiation potential could be maintained when expanded for more than 70 cell doublings by reconstitution of telomerase activity,which provided the novel experimental methods to establish the standard cell lines.
出处
《中国生物工程杂志》
CAS
CSCD
2004年第8期48-53,共6页
China Biotechnology
基金
国家杰出青年基金资助项目 ( 3 0 12 5 0 18)
国家"863"计划资助项目 ( 2 0 0 2AA2 0 5 0 61)
