猪骨髓间充质干细胞的分离培养及分化潜能的鉴定

Isolation and culture of multipotent mesenchymal stem cells from porcine bone marrow

目的:建立猪骨髓间充质干细胞(MSCs)的体外分离培养和鉴定的方法,探讨体外培养的间充质干细胞的一些生物学特点,为利用猪的实验研究提供实验基础。方法:从猪的髂嵴穿刺取骨髓,经密度梯度离心得到骨髓单个核细胞,接种后形成单层贴壁的成纤维样的细胞。检测细胞周期,多向诱导分化鉴定分离的细胞。结果:体外培养的原代MSCs 12～14d达到融合,传代后仍具有分化成骨的能力,细胞周期显示有80%细胞处于G_0/G_1期。结论:体外培养猪的MSCs具有分化成骨的潜能,生长稳定,传代后仍保持未分化状态,猪骨髓间充质干细胞分离培养体系的建立为基础研究和组织工程提供了一个有价值的动物模型。

To establish a method for the isolation of porcine msenchymal stem cells (MSCs) from bone marrow and to demonstrate their ex vivo differentiation into various mesenchymal tissue cells. Methods: MSCs were isolated from bone marrow and purified by centrifugation in vitro. The proliferation and growth characteris tics were observed in primary and passage cultures. Cell cycle was analyzed by measuring DNA content with flow cytometer and multipotent cells were identified by specific staining. Results:The adherent, fibroblast-shaped cells became confluent in a single layer on days 12 to 14 after plating. The cultured MSCs were differentiated into osteo blasts in vitro. The cell cycle analysis showed that 80% of MSCs were in G_0/G_1 phase. Conclusion: Porcine MSCs can be isolated from bone marrow by their adherent ability. Porcine MSCs may be recommended as a valuable model to study the mesenchymal lineages for basic research and tissue engineering,