Human embryonic stem cells (hESCs) have unique abilities that enable their use in cell therapy, disease modeling, and drug development. Their derivation is usually performed using a feeder layer, which is undefined and can potentially cause a contamination by xeno components, therefore there is a tendency to replace feeders with xeno-free defined substrates in recent years. Three hESC lines were successfully derived on the vitronectin with a truncated N-terminus (VTN-N) in combination with E-cadherin in xeno-free conditions for the first time, and their undifferentiated state, hESC morphology, and standard karyotypes together with their potential to differentiate into three germ layers were confirmed. These results support the conclusion that the VTN-N/E-cadherin is a suitable substrate for the xeno-free derivation of hESCs and can be used for the derivation of hESCs according to good manufacturing practices.

Department of Experimental Biology Faculty of Science Masaryk University Kamenice 5 625 00 Brno Czech Republic

Department of Gynecology and Obstetrics Faculty of Medicine Center of Assisted Reproduction Masaryk University Brno and University Hospital Obilni Trh 11 602 00 Brno Czech Republic

Department of Histology and Embryology Faculty of Medicine Masaryk University Kamenice 5 625 00 Brno Czech Republic

International Clinical Research Center Cell and Tissue Engineering Facility St Anne's University Hospital Pekarska 53 602 00 Brno Czech Republic

International Clinical Research Center Cell and Tissue Regeneration St Anne's University Hospital Pekarska 53 602 00 Brno Czech Republic

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