骨髓间充质干细胞与聚3-羟基丁酸酯-co-4-羟基丁酸酯生物材料共培养细胞补片的初步研究  

Pilot Study on Bone Marrow Mesenchymal Stem Cells Cultured with Biological Materials P(3HB-co-4HB) as Cell Patch

作　　者：牛红星[1] 穆军升[1] 张健群[1] 胡平[2] 伯平[1] 王艳[2] 

机构地区：[1]首都医科大学附属安贞医院,北京100029 [2]清华大学化工系高分子研究所,北京100084

出　　处：《中国生物工程杂志》2012年第10期74-79,共6页China Biotechnology

基　　金：北京市人事局2008年度留学人员科技活动择优资助基金(LXZZ2008011);北京市卫生系统高层次卫生技术人才培养(2011-3-065)资助项目

摘　　要：目的:研究聚3-羟基丁酸酯-co-4-羟基丁酸酯[P(3HB-co-4HB)]这种新型高分子材料与骨髓间充质干细胞(BMSCs)共培养,观察材料对干细胞的存活及增殖的影响,形成细胞补片的效果;从而找到一种适合BMSCs生长、增殖的高分子生物材料,作为治疗心肌梗死,软骨损伤等多种组织损伤疾病的修复方法之一。方法:取清洁级雄性健康BSL-C57小鼠作为实验对象,通过分离培养获得小鼠BMSCs,并进行流式细胞仪鉴定表面标志物。BMSCs培养至5代后,将BMSCs与P(3HB-co-4HB)制成的生物材料薄膜共培养,24h后固定进行电镜扫描,并用DAPI荧光染料染色处理,在荧光显微镜下观察并进行细胞计数,并描绘生长曲线。结果:BMSCs流式细胞术鉴定:CD34、CD45阴性,CD90弱阳性,CD73阳性。扫描电镜下,P(3HB-co-4HB)材料与BMSCs共培养形成的细胞补片,其表面细胞数量多,细胞状态正常。荧光显微镜下,对其细胞补片表面的细胞进行计数,并绘制生长曲线,显示表面细胞有逐渐增多的趋势。结论:P(3HB-co-4HB)材料与BMSCs共培养制成的细胞补片表面有细胞存活及增殖,由于P(3HB-co-4HB)材料本身具有良好的生物组织相容性及可降解等性质,所以该新型高分子材料可以作为干细胞治疗多种疾病的支架材料之一。Objective： To study poly 3-hydroxybutyrate-co-4-hydroxybutyrate [P（3HB-co-4HB）] new polymer materials co-cultured with bone marrow mesenchymal stem cells,observe the impact of materials of cell proliferation,find a polymer biomaterials fit for the stem cell survival,as one of the treatment of variety of diseases patch.Method：Take clean male healthy BSL-C57 mice as an experiment animals,use the 3rd generations through isolated and cultured for murinev and the identification with flow cytometry.Use 3rd generation bone marrow mesenchymal stem cell and P（3HB-co——4HB） made of biological material film a total of culture,After 24h fixed conducted scanning electron microscopy,and using DAPI fluorescent dye staining processing observed by fluorescence microscopy and cell count,and describe the counting growth curve.Results： The result of flow cytometry identification is CD34,CD45 negative,CD90 weakly positive,CD73 positive.The scanning electron microscope,at P（3HB-co-4HB） surface a large number of normal cell state.Draw the growth curve with cell counting.Mean that the surface of cells is a growing tendency.Conclusion： P（3HB-co-4HB） materials and BMSCs cultured made cell patch surface cell survival,proliferation,due to P（3HB-co-4HB）,the material itself has a good biological histocompatibility and biodegradable nature,became one of the good materials for cell therapy.

关 键 词：骨髓间充质干细胞 干细胞治疗 细胞补片 聚3-羟基丁酸酯-co-4-羟基丁酸酯 

分 类 号：Q819[生物学—生物工程]