骨膜提取物诱导小鼠骨髓间充质干细胞分化为类成骨细胞

Induction of mouse bone marrow derived stem cells into osteoblast-like cells by bone membrane extract

目的应用骨膜提取物诱导骨髓间充质干细胞(MSCs)分化为类成骨细胞,为骨组织工程提供充足的种子细胞。方法切取小鼠颅骨骨膜提取物,诱导小鼠骨髓间充质干细胞。诱导分化后的细胞采用ALP染色、RT-PCR、蛋白印迹技术进行检测。结果形态学观察显示,分化后的MSCs细胞形态从长梭形变成三角形、多角形或立方形;ALP活性结果显示,分化后的MSCs细胞ALP活性远高于未分化的MSCs;蛋白印迹显示,分化后的MSCs表达成骨细胞相关的BMP-2蛋白;RT-PCR结果表明,分化后的MSCs表达成骨细胞相关的4个基因Type(I)collagen、Osteocalcin、ALP和BMP-2,而未分化的MSCs则几乎不表达。结论骨膜提取物可以诱导小鼠骨髓间充质干细胞分化为类成骨细胞。

Objective To study the induction of mouse bone marrow derived mesenchymal stem cells（MSCs） into osteoblast-like cells applying bone membrane extract to provide sufficient seeding cells for bone tissue engineering. Methods ALP staining, RT-PCR and Western blot methods were utilized to characterize the osteoblast-like cells after transdifferentiation. Results The morphology of differentiated MSCs was changed from spindle shape into triangle, cub or polygon shape. ALP activity assay showed that ALP activity of transdifferentiated MSCs was higher than that of MSCs. The expression level of BMP-2 in bone membrane extract treated MSCs was higher than that of untreated MSCs, and also transdifferentiated MSCs expressed osteoblast cells associated four genes （Type（I） collagen, osteocalcin, ALP and BMP-2） by RT-PCR, but not in untreated MSCs. Conclusion Bone membrane extract as an inducer could induce mouse MSCs to transdifferentiate into osteoblast-like cells.