Established a Preimplantation Genetic Testing-derived and Xeno-free Human Embryonic Stem Cell Line / 中山大学学报(医学科学版)

ABSTRACT

@#【Objective】 A human embryonic stem cell line derived from Preimplantation genetic testing （PGT） embryos was established in a xeno- free stem cell culture system to provide disease models for medical research. 【Methods】The xeno-free culture system using xeno-free human foreskin fibroblast feeder layers（XF-HFF）mixed with commercially available chemically-defined medium（CDM）was assessed. In the culture system，a new hESC cell line was established using discarded embryos derived from PGT in patients with chromosomal balance translocation.【Results】The new availabled stem cell line was successfully cultured in the xeno-free culture system for a long time（> 45 passages）. The karyotype analysis revealed that the new line kept the same karyotype over 45 passages. Moreover，the expression of pluripotent markers was detected by fluorescent immunostaining including SSEA- 3，SSEA- 4，SSEA- 1，TRA- 1- 60， and TRA-1-81. RT-PCR analysis showed that the stem cell markers were present in hESC grown on XF-HFF-CDM. In addition，the teratoma formation analysis demonstrated that the cells cultured in XF-HFF/CDM maintained their pluripotency in vivo.【Conclusions】Our study may provide the possibility to establish embryonic stem cells with certain pathogenic genes，which could be applied for clinical research and treatment.