摘要目的 观察过表达Bmi1基因对人胚胎干细胞H1体外造血分化产生集落的影响.方法 人胚胎干细胞H1与小鼠骨髓间充质干细胞OP9共培养的方法体外模拟造血分化,在共培养第8天用磁珠分选筛选出CD34+造血前体细胞进行集落形成实验,诱导产生各系造血集落.用实时荧光定量PCR和Western印迹分别检测Bmi1基因和BMI1蛋白在人胚胎干细胞H1体外造血分化产生CD34+前体细胞过程中的表达变化.将过表达Bmi1基因的人胚胎干细胞H1用同样方法进行造血分化,比较其与正常的人胚胎干细胞H1体外造血分化产生集落的情况.结果 Bmi1基因在人胚胎干细胞H1体外造血分化产生CD34+前体细胞过程中的表达先升高后下降,过表达Bmi1基因的H1细胞相比正常H1细胞,集落形成实验产生红系集落和混合系集落的数目明显增加.结论 过表达Bmi1基因促进人胚胎干细胞H1体外造血分化过程中红系和混合系集落的产生.

abstractsObjective To study the effect of Bmil gene overexpression on hematopoietic differentiation colony-forming assay of human embryonic stem cells.Method Human embryonic stem cell H1 was co-cultured with mouse bone marrow mesenchymal stem cells OP9 to simulate hematopoietic differentiation in vitro.On the eighth day of co-culture,CD34 + hematopoietic progenitor cells were selected for colony-forming assay to induce hematopoietic colonies.The expression of Bmi1 gene and BMI1 protein in the process of H1 in vitro hematopoietic differentiation to CD34 + precursor cells was respectively detected by real-time PCR and Western Blot.The human embryonic stem cell H1 which overexpress Bmi1 gene was subjected to hematopoietic differentiation in the same way.Compare the colonies generated by hematopoietic differentiation of the two types of H1 cells.Results The expression of Bmi1 gene was first increased and then decreased in the process of in vitro hematopoietic differentiation to CD34 + hematopoietic progenitor cells of human embryonic stem cells.Colony-forming assay showed H1 cells which overexpress Bmi1 gene produced significantly more erythroid colony forming units and mixed colony forming units than normal H1 cells.Conclusion Overexpression of Bmi1 gene promote the production of erythroid and mixed colony forming units during hematopoietic differentiation of human embryonic stem cell H1.