犬骨髓血管内皮祖细胞的分离和诱导分化

目的 :分离并诱导犬骨髓内皮祖细胞分化为内皮细胞。并探讨不同培养条件、血清浓度和接种密度对分化的影响。方法 :密度梯度离心法分离骨髓单个核细胞 ,以不同密度分别接种于DMEM和EBM 2 +SingleQuots培养基中 ,在不同浓度胎牛血清条件下培养。随后鉴定细胞并比较各组分化率。结果 :接种密度较低时细胞无法成活 ,接种密度达 10 5/cm2 时细胞能表达内皮细胞标志并具有内皮细胞功能。 2 %和 2 0 %胎牛血清培养细胞 ,其分化率存在显著差异。结论 :犬骨髓能够分离出内皮祖细胞并能在体外分化为内皮细胞 ;接种密度过低对其分化不利 ;低浓度胎牛血清有利于细胞的分化。

Isolation and Differentiation of Vascular Endothelial Progenitor Cells from Canine Bone Marrow

Objective:To study endothelial progenitor cells from canine bo ne marrow differentiated into mature endothelial cells.And the influences of culture dens ity and fetal bovine serum(FBS) concentration on cell differentiation.Method:The mononuclear cells were isolated by the gradient dens ity centrifugation technique,cultured on the dish,supplied with the DMEM and E BM-2 added SingleQuots,and then incubated with different FBS concentr ation at 2% and 20%.The cell makers and functions were assayed by immunocytoche mistry and FACS Cailibur.Results:The cells cultured in low density are dead gradually, but the cells cultured in 10 5/cm 2 expressed the endothelial markers and func tions. The differentiation rates of cells cultured with 2% and 20% FBS are different significantly.Conclusion:The endothelial progenitor cells from bone marro w can be differentiated into endothelial cells.But it can not be succeeded if the culture density is too low,and the low FBS concentration is advantaged to keep the cells growing into endothelial cells.