拮抗TGFβ1可抑制胚胎干细胞来源BLCFC产生及分化.doc

拮抗TGFβ1可抑制胚胎干细胞来源BLCFC产生及分化 　 作者：要晖宇 刘兵 王晓燕 毛宁 论文代写 【摘要】 　　胚胎干细胞来源的BLCFC体外可分化产生造血和内皮细胞。为了 研究 TGFβ1对BLCFC产生和分化的调控作用，在拟胚体培养阶段施加TGFβ1和TGFβ1中和抗体，观察不同阶段TGFβ1对BLCFC的数目及其定向内皮和造血细胞分化能力的 影响 。结果表明：在拟胚体培养阶段拮抗TGFβ1的作用可显著降低BLCFC的数量(Plt;0.01)；与之吻合的，中和TGFβ1后拟胚体中Flk1+细胞比例降低。此外，在拟胚体培养阶段加入TGFβ1可显著增强BLCFC的造血和内皮细胞分化能力，而中和TGFβ1后分化能力降低。结论： TGFβ1对BLCFC的产生和分化发挥正向调节作用。 毕业论文 【关键词】 TGFβ1；BLCFC；胚胎干细胞 　　 Reduction of BlastColony Forming Cell Development and Bipotential Commitment by Neutralizing TGFβ1 In Vitro Abstract To study the regulation of TGFβ1 on the development of hemangioblast, embryonic stem cellderived blast forming cells (BLCFC) were used as the model of hemangioblast in vitro. TGFβ1 or antiTGFβ1 neutralization antibody was added in the medium of embryoid body (EB) generation for observating influence of TGFB1 addition in different culture stages on number of BLCFC and differentiation of BLCFC to endothelial and hematopoietic cells. The results showed that antagonizing TGFβ1 in the course of EB growth could significantly reduce the number of BLCFC (Plt;0.01), and the frequency of Flk1+ cells was also decreased consistently. Furthermore, the BLCFC derived from EB pretreated with TGFβ1 demonstrated remarkably elevated hematopoietic and endothelial potential, whereas such bipotential was impaired in the group with neutralizing antibody. It is concluded that TGFβ1, a conventional negative regulator in hematopoiesis and angiogenesis exert positive effects on the development and differentiation capacities of BLCFC in vitro. Key words TGFβ1； BLCFC； embryonic stem cells 胚胎干细胞(embryonic stem cells, ESC) 来源的BLCFC (blastcolony forming cell) 的检测是研究成血血管细胞(hemangioblast)的主要体外模型，已经为阐明某些转录因子和细胞因子在成血血管细胞发育中的调控作用提供了有效的途径［1,2］。普遍的观点认为，TGFβ1是成体造血和内皮细胞增殖的抑制因子；TGFβ1－/－小鼠卵黄囊造血和内皮系统的发育异常也表明其在胚胎早期发育中对于中胚层细胞分化、卵黄囊血管发生极为重要。此外，TGFβ1可体外诱导鸟类体壁中胚层细胞分化的同时产生造血和内皮细胞，提示其在成血血管细胞发育中具有重要作用。最近的研究证明，TGFβ1信号通路下游分子Smad5基因敲除的ESC中BLCFC数量明显高于野生型［3］。因此，TGF