蒙古马骨髓间充质干细胞的分离培养及多向分化潜能的研究

Study on Isolation,Culture and Multiple Differentiation Potential of Mongolia Horse Bone Marrow Derived Mesenchymal Stem Cells

旨在探索一个高效分离和扩增蒙古马骨髓间充质干细胞的方法,并通过形态学和多向分化能力进行鉴定。抽取蒙古马骨髓标本,利用密度梯度离心和差异贴壁法分离MSCs,体外扩增后传代,进行相差显微镜形态学观察。在地塞米松、IBMX、胰岛素及罗格列酮的作用下向脂肪细胞诱导分化,以及在地塞米松、VitC、β-磷酸甘油等作用下向成骨细胞诱导分化。结果显示原代和传代细胞呈梭形成纤维细胞样外观,生长增殖能力良好。经定向诱导分化后,细胞分别呈现脂肪细胞、成骨细胞的表型特征。证明所纯化的细胞为尚未分化的基质细胞,而不是组织的成体细胞,它具有多向分化潜能。

To establish a method for effective isolation and amplification of Mongolia horse bone-marrow derived mesenchymal stem cells（MSCs） and verification by morphology and multiple differentiation potential.MSCs were isolated from Mongolia horse bone marrow by gradient centrifugation and different adherent time method.Morphology and growth characteristics were examined by phase contrast microscopy.MSCs were induced in vitro to adipocytes by dexamethasone,IBMX,insulin and rosiglitazone,and to osteoblasts by β-glycerophosphate,L-ascorbicacid-2-phosphate and dexamethasone.The result showed that primary and passage cells were spindle-shaped and presented active proliferation.Cells were successfully induced to adipocytes and osteoblasts cells,and presented the characteristics of adipocytes and osteoblasts cells.The result indicate that the method is established to isolate and multiply MSCs from Mongolia horse bone marrow effectively and to verify by morphology and its multipotential differentiation.It prove that the purified cells are not adult somatic cells but matrix cells.