摘要
目的评估红系祖细胞在先天原始造血缺陷cloche~(-/-)突变体斑马鱼体内的移植效果。方法收集绿色荧光标记红系祖细胞的转基因系zTg(gata1:EGFP)斑马鱼胚胎,制备成单细胞悬液,经流式细胞仪分选出携带绿色荧光的gata1~+细胞,利用显微注射技术将gata1~+细胞移植到42hpf的cloche~(-/-)突变体斑马鱼心脏中,采用体视荧光显微镜追踪观察移植后的红系祖细胞在cloche~(-/-)突变体斑马鱼中的表达情况。结果成功分选出携带绿色荧光的gata1~+细胞,并在移植后2 h可观察到携带绿色荧光的gata1~+细胞逐渐增殖扩散且16 h持续有绿色荧光表达。结论红系祖细胞有重建造血的潜力,为进一步研究红系祖细胞移植后的功能鉴定提供实验依据。
Objective To evaluate the transplantation effect of erythroid progenitor cells in cloche~(-/-) mutant zebrafish with congenital primary hematopoietic deficiency. Methods The embryos of zebrafish zTg(gata1:EGFP) were collected and prepared into a single cell suspension. The gata1~+ cells carrying green fluorescence were separated by flow cytometry and transplanted into the hearts of 42 hpf cloche~(-/-) mutant zebrafish by microinjection. The expression of erythroid progenitor cells in cloche~(-/-) mutant zebrafish was examined by stereo fluorescence microscopy. Results gata1~+ cells with green fluorescence were successfully selected. The proliferation of gata1~+ cells with green fluorescence was observed at 2 h after transplantation, and 16 h of continuous green fluorescence expression was observed. Conclusions Erythroid progenitor cells have a potential to reconstruct hematopoiesis and provide an experimental basis for the further study of the functional identification of erythroid progenitor cells after transplantation.
引文
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