Human embryonic stem cells (hESCs) are increasingly used in clinical trials as they can change the outcome of treatment for many human diseases. They are used as a starting material for further differentiation into specific cell types and to achieve the desirable result of the cell therapy; thus, the quality of hESCs has to be taken into account. Therefore, current good manufacturing practice (cGMP) has to be implemented in the transport of embryos, derivation of inner cell mass to xeno-free, feeder-free and defined hESC culture, and cell freezing. The in-depth characterization of hESC lines focused on safety, pluripotency, differentiation potential and genetic background has to complement this process. In this paper, we show the derivation of three clinical-grade hESC lines, MUCG01, MUCG02, and MUCG03, following these criteria. We developed and validated the system for the manufacture of xeno-free and feeder-free clinical-grade hESC lines that present high-quality starting material suitable for cell therapy according to cGMP.

Cell and Tissue Engineering Facility International Clinical Research Center St Anne's University Hospital Pekarska 664 53 60200 Brno Czech Republic

Cell and Tissue Regeneration International Clinical Research Center St Anne's University Hospital Pekarska 664 53 60200 Brno Czech Republic

Center of Assisted Reproduction Department of Gynecology and Obstetrics University Hospital Brno Masaryk University Jihlavska 20 62500 Brno Czech Republic

Department of Experimental Biology Faculty of Science Masaryk University Kamenice 5 62500 Brno Czech Republic

Department of Histology and Embryology Faculty of Medicine Masaryk University Kamenice 5 62500 Brno Czech Republic

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