Gao Jiao, Yao Hui-Yu, Liang Xiao-Lei, Wang Xiao-Yan, Wu Ying, Liu Yuan-Lin, Mao Ning
Clinical Laboratory, PLA Hospital 306, Beijing, China.
Zhongguo Shi Yan Xue Ye Xue Za Zhi. 2011 Oct;19(5):1230-3.
This study was aimed to investigate whether endothelium-specific deletion of PTEN can affect hemangioblast development in the AGM region of mouse embryos. Based on Cre/loxP system, the Tie2CrePten(loxp/loxp) and Tie2CrePten(loxp/wt) mouse embryos were obtained. The genotype was identified by PCR. After treated with type I collagenase, the AGM region was dispersed into single-cell suspension, and then was cultured in blast colony-forming cell (BL-CFC) media. The number of BL-CFC was counted 4 or 5 days later. The hematopoietic capacity of BL-CFC was detected in methylcellulose culture system and the endothelial potential was assessed by tube-like structure formation on Matrigel. The results showed that the number of BL-CFC in AGM region of Tie2CrePten(loxp/loxp) mouse embryo decreased as compared with Tie2CrePten(loxp/wt) embryo. Whereas the hematopoietic capacity of mutant BL-CFC was enhanced, the endothelial potential, as evaluated by tube-like structure formation in vitro, was significantly reduced. It is concluded that the endothelial PTEN is capable of exerting regulatory functions on both the numbers and the dual potential of hemangioblast in mouse AGM region.
本研究旨在探讨内皮细胞特异性缺失PTEN是否会影响小鼠胚胎主动脉-性腺-中肾（AGM）区造血干细胞的发育。基于Cre/loxP系统，获得了Tie2CrePten(loxp/loxp)和Tie2CrePten(loxp/wt)小鼠胚胎。通过聚合酶链反应（PCR）鉴定基因型。用I型胶原酶处理后，将AGM区分散成单细胞悬液，然后在胚泡集落形成细胞（BL-CFC）培养基中培养。4或5天后计数BL-CFC的数量。在甲基纤维素培养系统中检测BL-CFC的造血能力，并通过在基质胶上形成管状结构来评估其内皮潜能。结果显示，与Tie2CrePten(loxp/wt)胚胎相比，Tie2CrePten(loxp/loxp)小鼠胚胎AGM区的BL-CFC数量减少。虽然突变型BL-CFC的造血能力增强，但通过体外形成管状结构评估的内皮潜能显著降低。结论是，内皮PTEN能够对小鼠AGM区造血干细胞的数量和双潜能发挥调节作用。
