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    • 10. 发明公开
    • METHOD FOR PRODUCING ENDOTHELIAL CELLS (VARIANTS)
    • VERFAHREN ZUR PRODUKTION VON ENDOTHELZELLEN(VARIANTEN)
    • EP2277993A1
    • 2011-01-26
    • EP08873385.2
    • 2008-12-24
    • Obshhestvo S Ogranichennojj Otvetstvennost' Ju "Laboratorija Kletochnykh Tekhnologijj"
    • KISELJOV, Sergejj L'vovichLAGAR'KOVA, Marija Andreevna
    • C12N5/08
    • C12N5/069C12N2500/90C12N2501/115C12N2501/125C12N2501/15C12N2501/155C12N2501/165C12N2506/02C12N2533/54
    • The invention relates to biotechnology, in particular to producing endothelial cells from human embryonic stem cells and to the use thereof. The inventive method for producing endothelial cells from human embryonic stem cells involves differentiating human embryonic stem cells on a substrate and in a synthetic medium that provide targeted direct differentiation of hESCs into endothelial cells based on a mixture of DMEM/F12 and a KO serum substitute having VEGF, SCF, BMP4, bFGF and TGFbeta growth factors and taken in effective quantities, wherein TGFbeta is introduced on 3rd-6th day of differentiation. A differentiation medium based on the mixture of DMEM/F12 with fetal bovine serum having VEGF, SCF and bFGF growth factors can be also used. The separation is carried out on 3rd-9th day according to an immunological selection method using markers specific for endothelial cells and, afterwards, the thus produced endothelial cells are cultivated in the abovementioned medium at the plating density of not less than 50,000 cells per cm 2 .
    • 本发明涉及生物技术,特别涉及从人类胚胎干细胞产生内皮细胞及其应用。 用于从人胚胎干细胞产生内皮细胞的本发明的方法包括在基质和合成培养基中分化人胚胎干细胞,所述培养基基于DMEM / F12和KO血清替代物的混合物将hESC靶向直接分化为内皮细胞,所述混合物具有 VEGF,SCF,BMP4,bFGF和TGFbeta生长因子,有效量,其中TGFbeta在分化第3〜6天引入。 还可以使用基于DMEM / F12与具有VEGF,SCF和bFGF生长因子的胎牛血清的混合物的分化培养基。 根据使用对内皮细胞特异性标记的免疫学选择方法,在第3〜9天进行分离,之后,将上述制作的内皮细胞以不小于50,000个细胞/ cm 2的电镀密度培养在上述培养基中 。