酸性成纤维细胞生长因子与胚胎干细胞的造血分化

背景：有研究表明，在卵黄囊造血、胎肝造血和胚胎干细胞向造血干细胞分化过程中，酸性成纤维细胞生长因子可强烈表达。目的：探讨酸性成纤维细胞生长因子对小鼠胚胎干细胞造血分化的作用，在拟胚体培养阶段施加酸性成纤维细胞生长因子，验证酸性成纤维细胞生长因子对造血形成细胞产生的调控作用。方法：培养小鼠胚胎干细胞，将饲养层上生长状态良好的小鼠胚胎干细胞用胰酶消化成单个细胞后，利用悬滴法制备拟胚体，拟胚体继续悬浮培养，以1，2和50g／L酸性成纤维细胞生长因子分别培养3，5，7，9d，通过免疫荧光法检测胚胎干细胞与拟胚体中酸性成纤维细胞生长因子的表达，流式细胞术检测FIk-1^＋与CD133^＋阳性细胞率。结果与结论：酸性成纤维细胞生长因子在拟胚体中呈阳性表达。在5μg／L酸性成纤维细胞生长因子作用下，FIk-1^＋细胞随时间增加表达增加，CD133^＋细胞表达模式与FIk-1^＋细胞类似。在1μg／L时，FIk-1^＋细胞在7d时表达达到高峰，随后下降。CD133^＋细胞表达结果类似。而在2pg／L时，5d时FIk-1^＋细胞表达升高，随后下降，在9d时表达又增高。而CD133^＋细胞则总体呈现升高趋势。酸性成纤维细胞生长因子可促进FIk-1^＋与CD133^＋细胞的产生，证明酸性成纤维细胞生长因子能够有效促进拟胚体的扩增以及成血管血液干细胞的产生与增殖。

Acidic fibroblast growth factor effect on hematopoietic differentiation of embryonic stem cells

BACKGROUND： Studies have shown that acidic fibroblast growth factor can be strongly expressed in the yolk sac hematopoietic cells and fetal liver hematopoietic cells and in the differentiation process of embryonic stem cells to hematopoietic stem cells. OBJECTIVE： To explore the effect of acidic fibroblast growth factor on hematopoietic differentiation of mice embryonic stem cells, to test and verify the regulation effect of acidic fibroblast growth factor in the formation of hematopoietic cells through adding the acidic fibroblast growth factor into the embryoid body culture stages.METHODS： The mouse embryonic stem cells were cultured and then digested into single cells using trypsin when they were in good conditions on the feeder layer. The embryoid bodies were prepared with hanging drop method and then continue suspension cultured for 3, 5, 7 and 9 days with 1, 2 and 5 pg/L acidic fibreblast growth factor. The expression of acidic fibroblast growth factor in embryonic stem cell and embryoid bodies was detected with immunofiuorescence, and the FIk-1^＋ and CD133＋ positive rates were detected with flow cytometry. RESULTS AND CONCLUSION： Acidic fibroblast growth factor showed positive expression in embryoid bodies. In the effect of 5 μg/L acidic fibroblast growth factor, FIk-1^＋ cells expression was increased with time increasing, the CD133＋ cells expression patterns were similar to FIk-1^＋ cells. In the effect of 1 μg/L acidic fibreblast growth factor, FIk-1^＋ cells expression peaked at 7 days and then decreased, and the CD133＋ cells showed similar expression patterns. But in the effect of 2 pg/L acidic fibroblast growth factor, FIk-1^＋ cells expression was increased at 5 days, then decreased, and increased again at 9 days; while CD133＋ cells expression emerged a rising trend totality. Acidic fibroblast growth factor can promote the generation of FIk-1^＋ and CD133＋ cells, suggesting that acidic fibroblast growth factor can effectively promote the amplification of embryonic body and the production and proliferation of hemangioblasts.