大鼠骨髓间充质干细胞在静电纺丝素/聚乳酸纳米纤维上的培养及成神经诱导  

Growth and Neural Differentiation of Bone Marrow Mesenchymal Stem Cells Cultured on Silk Fibroin Polylactic Acid Electrospun Nanofibers

作　　者：宋琳[1] 朱爱思[1] 徐路尧[1] 张锋[2] 左宝齐[2] 王敏[1] 包普花[1] 张焕相[1] 

机构地区：[1]苏州大学医学部江苏省干细胞研究重点实验室,江苏苏州215123 [2]苏州大学材料工程学院,江苏苏州215021

出　　处：《苏州大学学报（医学版）》2009年第3期396-399,F0002,共5页Suzhou University Journal of Medical Science

基　　金：国家重点基础研究发展计划(973计划)项目(2005CB623902);江苏省高校自然科学基金资助项目(05KJB310115);江苏省丝绸工程重点实验室;苏州大学医学发展基金资助项目(EE120602)

摘　　要：目的通过将骨髓间充质干细胞(BMSCs)培养在静电纺丝素/聚乳酸纳米纤维上,研究BMSCs的生长及成神经分化情况。方法用家蚕丝素、柞蚕丝素分别与聚乳酸共混制成静电纺丝素/聚乳酸纳米纤维,将第5代大鼠BMSCs培养其上,于24h后通过活细胞工作站观察细胞的黏附情况,并进行表型鉴定及存活检测。接种后待细胞长至60%左右,用bFGF/BHA诱导细胞成神经分化,并设多聚赖氨酸组进行对照。在诱导5h和维持48h时,观察细胞的形态学改变,通过免疫荧光法鉴定神经细胞特异性标志物Nestin,β-Ⅲ-Tubulin和NCAM的表达并进行定量统计分析。结果BMSCs在静电纺丝素/聚乳酸纳米纤维上的黏附情况良好,细胞生长于纳米纤维上。存活检测中几乎未发现死细胞,多数细胞在材料上可存活。神经分化的形态学改变与多聚赖氨酸组一致,并且培养在纳米纤维上的细胞分化时出现的突起可缠绕在纺丝纤维上。神经特异标志的表达情况与多聚赖氨酸对照组的差异无统计学意义(P>0.05)。结论静电纺丝素/聚乳酸纳米纤维具有良好的生物相容性,可支持BMSCs的黏附及成神经分化,且对细胞生存无毒性。Ojective To evaluate the growth and neural differentiation of bone marrow mesenchymal stem cells （BMSCs） cultured on the silk fibroin polyactic acid electrospun nano fibers. Methods In this study, the growth and neural differentiation of BMSCs on the electrospun nanofibers that were produced by using bombyx mori silk fibroin （B. mori SF）, antheraea pernyi silk fibroin （A. pernyi SFr） and PLA were analyzed. Results The results demonstrated that BMSCs adhered well and attached close to the nanofibers. Cells were positive for CD106, CD90 and negative for CD45 expression, phenotypic expressions characteristic of MSCs. In live and dead assay, no dead cells were found after cultured for 24 h. The nanofibers supported the growth of MSCs that underwent neurogenic differentiation as did poly-L-lysine which is commonly used for cell cultures. Conclusion Taken together, both two kinds of silk fibroin polylactic acid electrospun nanofibers are biocompatible and BMSCs can adhere and grow well on them. Also the nanofibers can support neural differentiation, and this could contribute to the development of tissue engineering applications for therapeutic uses.

关 键 词：静电纺丝素 聚乳酸纳米纤维 骨髓间充质干细胞 成神经分化 

分 类 号：R318.08[医药卫生—生物医学工程] R329.28[医药卫生—基础医学]