CAJ | 학술논문

背景：种子细胞的数量和质量是制约血管组织工程研究的重要瓶颈，而如何获取干细胞源内皮细胞是解决该瓶颈问题的关键。 　　目的：探讨添加血管内皮生长因子联合低氧环境是否能够协同高效诱导胚胎干细胞向内皮细胞定向分化。 　　方法：采用无血清培养基mTeSR?1维持培养人胚胎干细胞H9。通过添加血管内皮生长因子(50μg/L)联合低氧条件(体积分数为5%O2)诱导H9细胞向内皮细胞分化。采用免疫荧光染色技术、定量RT-PCR以及低密度脂蛋白摄取实验对人胚胎干细胞源内皮细胞进行表型和功能评价。 　　结果与结论：添加血管内皮生长因子联合低氧培养条件可促进H9细胞向内皮细胞方向分化。该胚胎干细胞源内皮细胞不但高水平表达内皮细胞的标志基因(kdr，pecam)和标记蛋白D31，而且还能够摄取低密度脂蛋白，形成类似微血管结构。提示血管内皮生长因子与低氧在诱导干细胞向内皮细胞分化过程中存在协同效应，可高效获得具有理想表型和功能活性的干细胞源内皮细胞。
배경：충자세포적수량화질량시제약혈관조직공정연구적중요병경，이여하획취간세포원내피세포시해결해병경문제적관건。 　　목적：탐토첨가혈관내피생장인자연합저양배경시부능구협동고효유도배태간세포향내피세포정향분화。 　　방법：채용무혈청배양기mTeSR?1유지배양인배태간세포H9。통과첨가혈관내피생장인자(50μg/L)연합저양조건(체적분수위5%O2)유도H9세포향내피세포분화。채용면역형광염색기술、정량RT-PCR이급저밀도지단백섭취실험대인배태간세포원내피세포진행표형화공능평개。 　　결과여결론：첨가혈관내피생장인자연합저양배양조건가촉진H9세포향내피세포방향분화。해배태간세포원내피세포불단고수평표체내피세포적표지기인(kdr，pecam)화표기단백D31，이차환능구섭취저밀도지단백，형성유사미혈관결구。제시혈관내피생장인자여저양재유도간세포향내피세포분화과정중존재협동효응，가고효획득구유이상표형화공능활성적간세포원내피세포。
BACKGROUND:The quantity and quality of seed cel s is a critical bottleneck of the development of vascular tissue engineering. To address this issue, stem cel-derived endothelial cel s have been a hot spot in this field due to their potential in providing the ideal seed cel s. OBJECTIVE:To elucidate the effect of vascular endothelial growth factor (VEGF) supplementation combined with hypoxic culture condition on the lineage-specific differentiation of embryonic stem cel s into endothelial cel s. METHODS:Serum-free medium mTeSR?1 was applied to cultivate H9 cel s in vitro. A conditioned medium containing 50μg/L vascular endothelial growth factor was utilized to induce H9 cel s to differentiate into endothelial cel s under the hypoxic culture condition (5%O2). The cel under normal condition (5%CO2) with or without vascular endothelial growth factor served as controls. The phenotype and function of human embryonic stem cel s-derived endothelial cel s were assayed by immunofluorescence staining, quantitative RT-PCR, and low-density lipoprotein uptake experiment. RESULTS AND CONCLUSION:Compared with the control group, the H9 cel s were induced to be differentiated into endothelial-like cel s more efficiently when they were cultivated under a conditioned medium with vascular endothelial growth factor supplementation under the hypoxic condition. These differentiated cel s not only expressed some important surface markers of endothelia cel s, including kdr, pecam, but also took in low-density lipoprotein to form microvessle-like structures. This culture system supports a synergy effect of vascular endothelial growth factor and hypoxic environment that can efficiently promotes the lineage-specific differentiation of embryonic stem cel s into endothelial cel s with good phenotype and functionality.