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骨髓间充质干细胞与3种生物材料共培养心肌补片的对比研究

Comparative study of bone marrow mesenchymal stem cells co-cultured with three kinds of biological materials for myocardial patch

作者: 牛红星  穆军升  张健群  胡平  伯平  王艳 
单位:首都医科大学附属北京安贞医院-北京市心肺血管疾病研究所(北京100029)
关键词: 心肌再生;骨髓间充质干细胞;心肌梗死;生物材料 
分类号:
出版年·卷·期(页码):2012·31·5(482-486)
摘要:

目的 寻找一种适于骨髓间充质干细胞(bone marrow mesenchymal stem cell, BMSC)生长的高分子生物材料,作为治疗心肌梗死的心肌补片。方法 取清洁级雄性健康C57小鼠,通过分离培养获得小鼠BMSC并进行流式细胞术鉴定,CD34和CD45阴性, CD90弱阳性,CD73阳性。细胞培养至5代后,将BMSC分别与聚氨酯(PU)、聚3-羟基丁酸酯-co-4-羟基丁酸酯P(3HB-co-4HB)和聚碳酸亚丙酯类(PC)制成的生物材料薄膜共培养(n=6),24 h后固定,电镜扫描,并用DAPI荧光染料染色处理,荧光显微镜下观察并进行细胞计数。结果 在扫描电镜下,PU材料和P(3HB-co-4HB)材料制成的补片表面细胞形态正常,数量较多,PC材料表面光滑,表面细胞数量极少,有细胞碎片等细胞裂解物。荧光显微镜下细胞计数,经SPSS12.0软件分析,PU组和P(3HB-co-4HB)组细胞计数无统计学差异(P>0.05),PC组与PU组,PC组与P(3HB-co-4HB)组细胞计数均有统计学差异(P<0.05)。结论 PU材料和P(3HB-co-4HB)材料制成的补片均有干细胞生长。P(3HB-co-4HB)具有良好的生物组织相容性及可降解等性质,是心肌补片更好的生物材料。

Objective  To find a polymer biomaterial suitable for bone marrow mesenchymal stem cell (BMSC) growth as a myocardial patch. Methods We obtain mouse BMSCs from clean grade healthy male C57 mice and identify the cells as CD34, CD45 negative, CD90 weakly positive and CD73 positive by flow cytometry. Cultured after five generations, BMSCs are co-cultured with polyurethane(PU), 3-hydroxybutyrate-co-4- hydroxybutyrate [P(3HB-co-4HB)] and polycarbonate class (PC) (n = 6) for 24 h, fixed for scanning electron microscopy and are stained with DAPI fluorescent dye, observed under fluorescence microscopy for cell count. Results The cell morphology of the PU patch and the P (3HB-co-4HB) patch are normal and the quantities of cells are large in the scanning electron microscope. The surface of PC patch is smooth with small quantity of BMSCs, cell debris and cell lysates. The difference of cell counts between PU group and P(3HB-co-4HB) group under the fluorescence microscope is not significant analyzed by one-way ANOVA with SPSS 12.0 software. The differences of cell counts between the PC group and the PU group, the PC group and the P(3HB-co-4HB) group are statistically significant (P<0.05). Conclusions PU material and P (3HB-co-4HB) material are both fit for BMSCs co-culture. With the nature of good tissue compatibility and biodegradability, P(3HB-co-4HB) is to be used as cell cardiomyoplasty surgery cardiac patch material.

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