CAJ | 학술논문

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5μg/L 酸性成纤维细胞生长因子分别培养3,5,7,9 d,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Flk-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,Flk-1+细胞随时间增加表达增加,CD133+细胞表达模式与 Flk-1+细胞类似.在1μg/L 时,Flk-1+细胞在7 d 时表达达到高峰,随后下降.CD133+细胞表达结果类似.而在2μg/L 时,5 d 时 Flk-1+细胞表达升高,随后下降,在9 d 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 Flk-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.
배경:유연구표명,재란황낭조혈、태간조혈화배태간세포향조혈간세포분화과정중,산성성섬유세포생장인자가강렬표체.목적:탐토산성성섬유세포생장인자대소서배태간세포조혈분화적작용,재의배체배양계단시가산성성섬유세포생장인자,험증산성성섬유세포생장인자대조혈형성세포산생적조공작용.방법:배양소서배태간세포,장사양층상생장상태량호적소서배태간세포용이매소화성단개세포후,이용현적법제비의배체,의배체계속현부배양,이1,2화5μg/L 산성성섬유세포생장인자분별배양3,5,7,9 d,통과면역형광법검측배태간세포여의배체중산성성섬유세포생장인자적표체,류식세포술검측Flk-1+여 CD133+양성세포솔.결과여결론:산성성섬유세포생장인자재의배체중정양성표체.재5μg/L 산성성섬유세포생장인자작용하,Flk-1+세포수시간증가표체증가,CD133+세포표체모식여 Flk-1+세포유사.재1μg/L 시,Flk-1+세포재7 d 시표체체도고봉,수후하강.CD133+세포표체결과유사.이재2μg/L 시,5 d 시 Flk-1+세포표체승고,수후하강,재9 d 시표체우증고.이 CD133+세포칙총체정현승고추세.산성성섬유세포생장인자가촉진 Flk-1+여 CD133+세포적산생,증명산성성섬유세포생장인자능구유효촉진의배체적확증이급성혈관혈액간세포적산생여증식.
@@@@BACKGROUND: Studies have shown that acidic fibroblast growth factor can be strongly expressed in the yolk sac hematopoietic cel s and fetal liver hematopoietic cel s and in the differentiation process of embryonic stem cel s to hematopoietic stem cel s. OBJECTIVE: To explore the effect of acidic fibroblast growth factor on hematopoietic differentiation of mice embryonic stem cel s, to test and verify the regulation effect of acidic fibroblast growth factor in the formation of hematopoietic cel s through adding the acidic fibroblast growth factor into the embryoid body culture stages. METHODS: The mouse embryonic stem cel s were cultured and then digested into single cel s 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 μg/L acidic fibroblast growth factor. The expression of acidic fibroblast growth factor in embryonic stem cel and embryoid bodies was detected with immunofluorescence, and the Flk-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, Flk-1+ cel s expression was increased with time increasing, the CD133+ cel s expression patterns were similar to Flk-1+ cel s. In the effect of 1 μg/L acidic fibroblast growth factor, Flk-1+ cel s expression peaked at 7 days and then decreased, and the CD133+ cel s showed similar expression patterns. But in the effect of 2 μg/L acidic fibroblast growth factor, Flk-1+ cel s expression was increased at 5 days, then decreased, and increased again at 9 days; while CD133+ cel s expression emerged a rising trend totality. Acidic fibroblast growth factor can promote the generation of Flk-1+ and CD133+ cel s, suggesting that acidic fibroblast growth factor can effectively promote the amplification of embryonic body and the production and proliferation of hemangioblasts.