胸腺肽类药物对体外诱导ES细胞分化为T细胞的作用_医学论文.docVIP

胸腺肽类药物对体外诱导ES细胞分化为T细胞的作用_医学论文 胸腺肽类药物对体外诱导ES细胞分化为T细胞的作用_医学论文 作者：彭延文， 徐霖， 张秀明， 张嘉晴， 李树浓， 项鹏 【摘要】 【目的】 研究目前临床常用的增强细胞免疫功能的胸腺肽，胸腺5肽和胸腺素α-1在小鼠胚胎干细胞向T淋巴细胞分化过程中的作用。【方法】 借助小鼠胚胎干细胞(ESC)体外自然分化形成的类胚体（EB）中含有三胚层细胞的独特细胞环境，加入多种细胞因子和胸腺多肽，体外诱导小鼠ESC向T淋巴细胞分化。利用流式细胞仪检测在三种不同胸腺多肽的诱导下，不同时间点的小鼠ESC来源的细胞表面CD3分子的表达水平。并在相应时间点通过RT-PCR检测与T淋巴细胞发育密切相关的Notch信号分子的转录水平。【结果】 在加入胸腺肽和胸腺素α1诱导的实验组，均有细胞表达CD3分子，CD3+细胞的百分比随诱导时间的增加而增多；而加入胸腺五肽的实验组无CD3+细胞出现。 加入胸腺素α1和胸腺肽的实验组，有Notch1及其配体delta-like-1和delta-like-4的转录；而加入胸腺五肽的实验组的细胞无Notch1及其配体转录。【结论】 胸腺素α1或胸腺肽可能通过影响Notch1信号途径支持ES细胞向T细胞分化，而胸腺五肽无此作用。 【关键词】 胚胎干细胞； T淋巴细胞； 胸腺5肽； 胸腺素α1； 胸腺肽 Abstract：【Objective】 To compare the effects of three thymic polypeptides including thymopentin-5 (TP5), thymosin-alpha1 (Tα-1) and thymopeptides on the thymopoiesis of mouse embryonic stem (ES) cells in vitro. 【Method】 EBs were obtained from spontaneously differentiated ES cells and were induced into T lymphocytes in vitro by utilizing three different inducing media, which contain TP5, Tα-1, and thymopeptides, respectively. Then the expression of CD3 on these differentiated cells were analyzed by flow cytometry at different time points. At the corresponding time-point, the transcription levels of Notch signal molecules were also detected by RT-PCR. 【Results】 The results showed that inducing systems containing either Tα-1 or thymopeptides promoted the emergence of CD3+ cells during the observation period, and the percentage of CD3+ cells increased with a longer induction period. However, we never found CD3+ cells in the inducing system containing TP5 throughout the inducing period. And Notch 1 receptor and its ligands, involving Delta-like-1 and Delta-like-4, expressed in inducing systems containing either Tα-1 or thymopeptides. However, neither Notch1 nor its ligands were detected in the inducing system containing TP-5 during the observation period. 【Conclusion】 Tα-1 and thymopeptides may play a role in promoting T cells differentiation from ES cells by Notch1 signal pathway, while TP