李方国;王磊;张鑫鑫;姜晓锐;金丹;江汕;赵培冉;裴国献;

• 1:南方医科大学南方医院创伤骨科
• 2:山东省烟台毓璜顶医院手足骨科
• 3:第四军医大学西京医院骨科

摘要(Abstract)：

[目的]探讨三种不同来源血清对体外培养成人骨髓基质干细胞(hBMSCs)向成骨诱导分化的作用。[方法]将体外第3代hBMSCs向成骨诱导分化培养,分为胎牛血清组(对照组)、AB血清组和自体血清组。对比观察细胞碱性磷酸酶(ALP)染色、钙结节染色。诱导培养后4、7、14 d和21 d,各血清组分别进行钙黄绿素法荧光显微镜动态观察矿盐沉积,检测ALP活性,实时荧光定量PCR(RT-qPCR)法检测成骨基因(ALP)、骨桥蛋白(OPN)和骨钙素(OCN)的表达。[结果]ALP染色和钙结节染色结果显示,与自体血清(AS)组、胎牛血清(FBS)组相比,AB血清(ABS)组明显提高了hBMSCs的染色阳性率。荧光显微镜下观察诱导21 d的hBMSCs,ABS组较FBS组、AS组呈现更多的钙盐沉积。ALP活性结果显示,同一时间点ABS组的ALP活性均明显高于FBS组和AS组,差异有统计意义(P<0.05)。RT-qPCR结果显示,在7、14 d和21d,ABS组ALP、OPN和OCN成骨基因表达均明显高于FBS组、AS组,其中,ALP基因表达在7 d出现峰值,OPN基因表达在14 d出现峰值,OCN基因表达在21 d出现峰值,差异均有统计学意义(P<0.05)。[结论]ABS对hBMSCs成骨分化作用较FBS、AS明显增强。ABS有望替代FBS建立符合骨组织工程临床应用要求的体外hBMSCs培养体系。

关键词(KeyWords)： 骨髓基质干细胞;AB血清;自体血清;成骨分化

Abstract:

Keywords:

基金项目(Foundation): 国家重点基础研究发展计划(973)资助项目(2009CB930003);; 国家自然科学基金项目(No.31070866)

作者(Author): 李方国;王磊;张鑫鑫;姜晓锐;金丹;江汕;赵培冉;裴国献;

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DOI:

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