组织工程材料捕获成体干细胞的影响因素研究  

Study on impact factors of tissue engineering materials capturing adult stem cell

作　　者：丁宁[1] 王世立[1] 韩金祥[1] 张翠[1] 葛鲁娜[1] 

机构地区：[1]山东省医学科学院/山东省医药生物技术研究中心/国家卫生部生物技术药物重点实验室/山东省医学分子生物学重点实验室,济南250062

出　　处：《中国医药生物技术》2009年第3期171-176,F0001,共7页Chinese Medicinal Biotechnology

基　　金：国家自然科学基金(30571766);山东省自然科学基金(Y2008C06)

摘　　要：目的探索体内埋植组织工程材料方法获取成体干细胞的相关影响因素。方法BALB/c小鼠随机分为2组,分别埋植经环氧乙烷灭菌的明胶海绵与聚乳酸,在植入后第3、7、14天时分别取出材料,HE染色观察细胞捕获情况。将埋植相同质量明胶海绵的小鼠随机分为成骨蛋白-1(OP-1)组、粒细胞集落刺激因子(G-CSF)组和对照组,在植入后第3、6、9、12、15天时分别取出材料,称取质量并进行细胞计数,采用流式细胞术检测第12天时分离获得的各组细胞表面CD34和干细胞抗原-1(Sca-1)的阳性表达率。另取第12天时分离获得的对照组细胞,分为OP-1组、G-CSF组和空白对照组,采用噻唑蓝(MTT)法检测细胞增殖活性。结果HE染色显示,明胶海绵与聚乳酸捕获的细胞数量均随时间延长而增加。各组植入的明胶海绵质量均随时间延长而减少,而捕获细胞数量均随时间延长而增加,OP-1组第9、12、15天和G-CSF组第9天时捕获细胞数量分别与对照组相比均有统计学意义(均P<0.05);OP-1组细胞表面CD34、Sca-1单阳性表达率和双阳性表达率(分别为13.07%±0.19%、3.98%±0.15%、17.02%±0.12%)均明显高于对照组(分别为11.62%±0.44%、3.03%±0.11%、2.91%±0.14%,均P<0.05),G-CSF组细胞表面CD34、Sca-1单阳性表达率(分别为12.79%±0.39%、4.52%±0.35%)分别与对照组相比均无统计学意义,但双阳性表达率(10.21%±0.15%)与对照组相比有统计学意义(P<0.05)。MTT检测结果显示,OP-1组和G-CSF组各时间点测定的吸光度值均高于空白对照组(均P<0.05)。结论通过体内埋植复合OP-1明胶海绵的方法可增加干细胞的捕获数量,为建立一种简易提取自体干细胞的新方法奠定了基础。Objective To explore the relevant factors of adult stem cells captured by implanting tissue engineering materials in vivo. Methods BALB/c mice were divided into 2 groups randomly, and respectively implanted absorbable gelatin sponge （AGS） and polymer of lactic acid （PLA） sterilized through ethylene oxide separately in vivo. The AGS and PLA were taken out and observed the captured cells in HE staining on days 3, 7, and 14. BALB/c mice implanted with the same quality of AGS were divided into osteogenic protein-1 （OP-1） group, granulocyte colony-stimulating factor （G-CSF） group, and the control group randomly. The AGS were taken out and weighed, then counted the number of the captured cells on days 3, 6, 9, 12, and 15. The positive rates of CD34 and stem cell antigen-1 （Sca-1） expression on the cells collected on day 12 in each group were detected by flow cytometry. The cells collected on day 12 in the control group were divided into OP-1 group, G-CSF group, and blank control group, and the proliferative activity of the captured cells was detected by MTT.Results HE staining showed that the number of the cells captured by AGS and PLA increased with time prolonged. In each group, the quality of AGS decreased with time prolonged, while the number of the captured cells increased with time prolonged. Compared with the control group, the number of the captured cells in the OP-1 group on days 9, 12, 15 and in the G-CSF group on days 9 were statistically significant （all P 〈 0.05）. Compared with the control group （11.62% ± 0.44%, 3.03% ± 0.11%, and 2.91% ± 0.14% respectively）, the single-positive and dual-positive rates of CD34 and Sca-1 expression （13.07% ± 0.19%, 3.98% ± 0.15 %, and 17.02% ± 0.12%, respectively） in the OP-1 group were significantly higher （all P 〈 0.05）, and the single-positive rates of CD34 and Sca-1 expression （12.79% ± 0.39% and 4.52% ± 0.35%） in the G-CSF group were not statistically significant, but the dual-positive rates （10.21% ± 0.15%） were

关 键 词：成体干细胞 明胶海绵 吸收性 骨形态发生蛋白质类 粒细胞集落刺激因子 

分 类 号：R318.08[医药卫生—生物医学工程] R329.2[医药卫生—基础医学]