FIELD: biotechnology.
SUBSTANCE: invention refers to biotechnology, namely to production of three-dimensional cell clusters and their differentiation. Method involves treatment of pluripotent stem cells, which are induced pluripotent stem cells, cells obtained from human umbilical cord tissue, parthenotes, cells obtained from amniotic fluid, or human embryonic stem cells of line H1, H7, H9, SA002 or BG01v, cultured in a flat adhesive culture together with a chelating agent or enzyme, with release of cell aggregates from a flat adhesive culture. That is followed by suspending cell aggregates from a flat adhesive culture into a culture medium in the presence of a Rho-kinase inhibitor without dissociating cell aggregates to single cells. Then, suspension of cell aggregates is transferred into dynamic suspension culture and increased volume of suspension of cell aggregates in dynamic suspension culture to obtain three-dimensional cell clusters, wherein three-dimensional cell clusters preserve pluripotency. Further, it is possible to differentiate pluripotent cell clusters in a dynamic suspension culture system to obtain a population of precursor cells of pancreatic cells, a population of neuronal progenitor cells or a population of cardiomyocyte precursor cells.
EFFECT: invention widens the range of agents.
34 cl, 20 tbl, 95 dwg, 16 ex
