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目的:从鼠胚胎干细胞获得具有造血重建功能的造血干细胞。方法:将ES-D3细胞形成4天拟胚体(4dEBs),再用骨髓内皮细胞条件培养液(mBMEC-CM)和/或胎肝基质细胞条件培养液(FLSC-CM)诱导4dEBs生成造血干细胞。实验为mBMEC-CM+FLSC-CM组、mBMEC-CM组、FLSC-CM组,对照组。检测诱导生成的细胞造血干细胞特异抗原表达、造血相关基因表达、造血集落的形成以及造血重建能力。结果:从诱导ES-D3细胞生成造血干/祖细胞的数量和生成的集落总数看,mBMEC-CM+FLSC-CM组诱导效率显著高于其他3组。mBMEC-CM+FLSC-CM组诱导生成的细胞能重建照射鼠的造血系统。结论:骨髓内皮细胞条件培养液联合胎肝基质细胞条件培养液可促进鼠胚胎干细胞分化为具有造血重建能力的造血干细胞。
Objective: To obtain hematopoietic stem cells with hematopoietic reconstitution function from mouse embryonic stem cells. METHODS: ES-D3 cells were cultured on embryonic day 4 (4dEBs) and induced by 4-day EBs (mBMEC-CM) and / or fetal liver stromal cell conditioned medium (FLSC-CM) . The experiments were mBMEC-CM + FLSC-CM group, mBMEC-CM group, FLSC-CM group and control group. The expression of hematopoietic stem cell-specific antigen, hematopoietic related gene expression, hematopoietic colony formation and hematopoietic reconstitution ability were detected. Results: The induced efficiency of mBMEC-CM + FLSC-CM group was significantly higher than that of the other three groups in terms of the number of induced hematopoietic stem / progenitor cells and the total number of colonies formed. The cells induced by mBMEC-CM + FLSC-CM group reconstituted the hematopoietic system of irradiated mice. CONCLUSION: Bone marrow endothelial cell conditioned medium combined with fetal liver stromal cell conditioned medium can promote the differentiation of mouse embryonic stem cells into hematopoietic stem cells capable of hematopoietic reconstitution.