Derivation and characterization of human embryonic stem cells on human amnion epithelial cells

• Published in: Scientific reports Vol. 5; no. 1; p. 10014
• Main Authors: Lai, Dongmei, Wang, Yongwei, Sun, Jian, Chen, Yifei, Li, Ting, Wu, Yi, Guo, Lihe, Wei, Chunsheng
• Format: Journal Article
• Language: English
• Published: London Nature Publishing Group UK 07.05.2015; Nature Publishing Group
• Subjects: 13; 13/1; 13/100; 13/106; 13/51; 14; 14/105; 14/28; 45; 631/532; 631/532/2117; Amnion; Amnion - cytology; Biomarkers; Blastocyst - cytology; Cell culture; Cell Culture Techniques; Cell Differentiation; Cell Line; Cell lines; Chromosome Banding; Cytoplasm; Electron microscopy; Embryo cells; Embryo fibroblasts; Embryonic Stem Cells - cytology; Embryonic Stem Cells - metabolism; Embryonic Stem Cells - ultrastructure; Embryos; Epithelial cells; Feeder Cells; Gene Expression Profiling; Gene Expression Regulation, Developmental; Humanities and Social Sciences; Humans; In vitro fertilization; multidisciplinary; Pluripotency; Pluripotent Stem Cells - cytology; Pluripotent Stem Cells - metabolism; Pluripotent Stem Cells - ultrastructure; Science; Stem cell transplantation; Stem cells; Teratoma; Transmission electron microscopy
• ISSN: 2045-2322; 2045-2322
• DOI: 10.1038/srep10014

Culture conditions that support the growth of undifferentiated human embryonic stem cells (hESCs) have already been established using primary human amnion epithelial cells (hAECs) as an alternative to traditional mitotically inactivated mouse embryonic fibroblasts (MEFs). In the present work, inner cell masses (ICM) were isolated from frozen embryos obtained as donations from couples undergoing in vitro fertilization (IVF) treatment and four new hESC lines were derived using hAECs as feeder cells. This feeder system was able to support continuous growth of what were, according to their domed shape and markers, undifferentiated naïve-like hESCs. Their pluripotent potential were also demonstrated by embryoid bodies developing to the expected three germ layers in vitro and the productions of teratoma in vivo . The cell lines retained their karyotypic integrity for over 35 passages. Transmission electron microscopy (TEM) indicated that these newly derived hESCs consisted mostly of undifferentiated cells with large nuclei and scanty cytoplasm. The new hESCs cultured on hAECs showed distinct undifferentiated characteristics in comparison to hESCs of the same passage maintained on MEFs. This type of optimized culture system may provide a useful platform for establishing clinical-grade hESCs and assessing the undifferentiated potential of hESCs.