Established a Preimplantation Genetic Testing-derived and Xeno-free Human Embryonic Stem Cell Line
- VernacularTitle:建立植入前遗传学检测人胚胎来源的无动物源性干细胞系
- Author:
Dan ZHANG
1
;
Zeng-yan WANG
1
;
Qing-yun MAI
1
;
Bing CAI
1
;
Yan-hong ZENG
1
;
Can-quan ZHOU
1
Author Information
1. Reproductive Medicine Center，The First Affiliated Hospital// Guangdong Provincial Key Laboratory of Reproductive Medicine；Sun Yat-sen University，Guangzhou 510080，China
- Publication Type:Journal Article
- Keywords:
human embryonic stem cells, xeno-free, PGT-derived
- From:
Journal of Sun Yat-sen University(Medical Sciences)
2019;40(4):510-517
- CountryChina
- Language:Chinese
-
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.
