FIELD: biotechnology.
SUBSTANCE: invention refers to biotechnology, namely to differentiation of a cellular cluster of pluripotent stem cells into definitive endoderm cells. Method involves producing a cellular cluster of pluripotent stem cells from pluripotent stem cells grown on a flat attached culture, formed from a cell cluster or not recovered from individual cell suspensions, wherein the pluripotent stem cells are non-embryonic pluripotent stem cells, inducible pluripotent cells, reprogrammed pluripotent cells, cells recovered from adult somatic cells, induced pluripotent stem cells, cells, obtained from human amniotic fluid, human parthenotes or cells from human embryonic stem cell lines H1, H7, H9 or SA002. Then, a cell cluster of pluripotent stem cells is transferred from a flat attached culture to a dynamic suspension culture without dissociating the cell cluster into separate cells and culturing a cellular cluster of pluripotent stem cells in a dynamic suspension culture in a culture medium, supplemented with (i) 14-prop-2-en-1-yl-3,5,7,14,17,23,27-hepta-tetra-tetracyclo[19.3.1.1~2,6-~.1~8,12.~]heptacosa-1(25),2(27),3,5,8(26),9,11,21,23-nonane-16-one and GDF8 or (ii) WNT3A and activator A or treatment of a pluripotent stem cell cell cluster in a dynamic suspension culture with said substances.
EFFECT: invention enables successful differentiation of a cellular cluster of pluripotent stem cells into definitive endoderm cells.
18 cl, 138 dwg, 16 ex, 19 tbl
