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Human embryonic stem cell (hESС) lines represent a cell population that has unlimited replicative capacity and can differentiate into all somatic cells and germ line. These properties makes hESC lines an excellent tool for human cell and tissue replacement therapy. In our work a novel hESC subline SC6-FF was derived in allogenic feeder-free culture conditions from hESC line SC6, which was derived with using of human feeder cells. The used culture system consists of extracellular matrix proteins and conditioned medium, synthesized by feeder cells – mesenchymal stem cell line SC5-MSC, which was previously derived from initial hESC line SC5.