脐带间充质干细胞条件培养液促进人胚胎干细胞分化为造血细胞

Human umbilical cord mesenchymal stem cells conditioned medium promotes the hematopoietic differentiation of human embryonic stem cells

目的利用人脐带间充质干细胞造血支持的特性,优化人胚胎干细胞造血分化方案。方法利用组织块培养法分离扩增人脐带间充质干细胞,收集条件培养液。以小鼠胚胎成纤维细胞为饲养层培养人胚胎干细胞,再用拟胚体培养液诱导拟胚体形成。分别用条件培养液和拟胚体培养液培养拟胚体,再用流式细胞术检测EB中造血标志CD45和CD34的表达。结果利用组织块培养法获得大量形态均一、增殖迅速的脐带间充质干细胞;间充质干细胞条件培养液诱导8d后,拟胚体中的CD45表达60%以上,CD34表达30%以上,远高于含有BMP4的拟胚体培养液的诱导率。结论人脐带间充质干细胞条件培养液可促进人胚胎干细胞的造血分化,并避免动物源污染,节约成本。

Objective To optimize the scheme of human embryonic stem cell differentiation to hematopoietic cell by taking advantage of the characteristics of umbilical cord mesenchymal stem cells in supporting hematopoiesis . Method The umbilical cord mesenchymal stem cells were isolated and expanded with tissue block culture method and then the condition medium were collected routinely .Human embryonic stem cell were cultured on mouse embryonic fibroblasts ,and were induced to be embryoid bodies with embryoid bodies culture medium .The embryoid bodies were cultured with condition medium and embryoid bodies culture medium respectively .Finally ,the percentage of hematopoietic cell maker CD45 and CD34 were detected with flow cytometry .Result It is obtained a large number of homogeneous fibroblasts‐like umbilical cord mesenchymal stem cells which proliferated rapidly .After the embryoid bodies were cultured with condition medium 8 days ,the result shown that the percentage of CD45 were more than 60% and CD34 were more than 30% ,far higher than that of embryoidbodies cultured with embryoid bodies culture medium .Conclusion Human umbilical cord mesenchymal stem cells conditioned medium can promote thehematopoiet‐ic differentiation of human embryonic stem cells .And his method will avoid the mouse‐related disease and cut down the cost of experiment .