Maintenance of human embryonic stem cells on gelatin

• Published in: Chinese science bulletin 2009-11, Vol.54 (22), p.4214-4220
• Main Authors: Li, Yang, Lin, ChangSheng, Wang, Li, Liu, Ying, Mu, XiaoNing, Ma, Yue, Li, LingSong
• Format: Article
• Language: English
• Subjects: Chemistry/Food Science; Earth Sciences; Engineering; Humanities and Social Sciences; Life Sciences; multidisciplinary; Physics; Science; Science (multidisciplinary); 人类胚胎干细胞; 培养体系; 涂层材料; 胰蛋白酶; 阳性克隆; 阳性细胞
• ISSN: 1001-6538; 1861-9541
• DOI: 10.1007/s11434-009-0630-4

Matrigel is routinely used as a coating material in the feeder-free culture system of human embryonic stem cells （hESCs）. However, matrigel is costive and inconvenient to use. In this study, the possibility of using gelatin as an alternative coating material was investigated. The results showed that, after trypsinization, hESCs were maintained undifferentiated on gelatin. These hESCs expressed pluripotent markers, formed teratoma and maintained a normal karyotype. As measured at passage 10, the hESCs expressed a high level of Oct4 on both gelatin and Matrigeh hESCs growing on gelatin formed AP-positive colonies in similar size and number to those growing on Matrigel （P〉 0.05）. Moreover, hESCs growing on gelatin contained a comparable percentage of SSEA-4-positive cells to those growing on Matrigel （95.1% vs.94.3%, P〉 0.05）. H-1 hESCs were maintained undifferentiated on gelatin for 20 passages and remained the stable normal karyotype. This gelatin-based culture protocol may allow us to propagate hESCs in large scale, with less cost.