A feeder-free culture using autogeneic conditioned medium for undifferentiated growth of human embryonic stem cells: Comparative expression profiles of mRNAs, microRNAs and proteins among different feeders and conditioned media

BMC Cell Biology

Research Article

Chi-Hsien Chou¹  Sung-Liang Yu²  Sher Singh³  Zong-Yun Tsai⁴  Steven Shoei-Lung Li⁵ 

[1] Center for Resources, Research and Development, Kaohsiung Medical University, 807, Kaohsiung, Taiwan;Department of Clinical Laboratory Sciences and Medical Biotechnology, National Taiwan University College of Medicine, 100, Taipei, Taiwan;Department of Life Science, College of Science, National Taiwan Normal University, 116, Taipei, Taiwan;Stem Cell Laboratory, Center of Excellence for Environmental Medicine, Kaohsiung Medical University, 807, Kaohsiung, Taiwan;Stem Cell Laboratory, Center of Excellence for Environmental Medicine, Kaohsiung Medical University, 807, Kaohsiung, Taiwan;Graduate Institute of Clinical Medicine, College of Medicine, Kaohsiung Medical University, 807, Kaohsiung, Taiwan;Correspondence to: Steven Shoei-Lung Li at Graduate Institute of Clinical Medicine, College of Medicine, Kaohsiung Medical University, 807, Kaohsiung, Taiwan;

关键词: Protein Spot;    Activin;    Mouse Embryonic Fibroblast;    Affymetrix GeneChip;    Process Network;   

DOI  :  10.1186/1471-2121-11-76

 received in 2010-01-15, accepted in 2010-10-12,  发布年份 2010

来源: Springer

【 摘 要 】

BackgroundHuman embryonic stem (hES) cell lines were derived from the inner cell mass of human blastocysts, and were cultured on mouse embryonic fibroblast (MEF) feeder to maintain undifferentiated growth, extensive renewal capacity, and pluripotency. The hES-T3 cell line with normal female karyotype was previously used to differentiate into autogeneic fibroblast-like cells (T3HDF) as feeder to support the undifferentiated growth of hES-T3 cells (T3/HDF) for 14 passages.ResultsA feeder-free culture on Matrigel in hES medium conditioned by the autogeneic feeder cells (T3HDF) was established to maintain the undifferentiated growth of hES-T3 cells (T3/CMHDF) for 8 passages in this investigation. The gene expression profiles of mRNAs, microRNAs and proteins between the undifferentiated T3/HDF and T3/CMHDF cells were shown to be very similar, and their expression profiles were also found to be similar to those of T3/MEF and T3/CMMEF cells grown on MEF feeder and feeder-free Matrigel in MEF-conditioned medium, respectively. The undifferentiated state of T3/HDF and T3/CMHDF as well as T3/MEF andT3/CMMEF cells was evidenced by the very high expression levels of "stemness" genes and low expression levels of differentiation markers of ectoderm, mesoderm and endoderm in addition to the strong staining of OCT4 and NANOG.ConclusionThe T3HDF feeder and T3HDF-conditioned medium were able to support the undifferentiated growth of hES cells, and they would be useful for drug development and toxicity testing in addition to the reduced risks of xenogeneic pathogens when used for medical applications such as cell therapies.

【 授权许可】

Unknown   
© Tsai et al; licensee BioMed Central Ltd. 2010. This article is published under license to BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.