Human Embryonic Stem Cells (hESCs) Cultured Under Distinctive Feeder-Free Culture Conditions Display Global Gene Expression Patterns Similar to hESCs from Feeder-Dependent Culture Conditions Tae-Min Yoon & Bomi Chang & Hyeung-Taek Kim & Joo-Hyun Jee & Dong-Wook Kim & Dong-Youn Hwang Published online: 3 June 2010 # Springer Science+Business Media, LLC 2010 Abstract Human embryonic stem cell (hESC)–based assay systems and genetically modified hESCs are very useful tools for screening drugs that regulate stemness and differentiation and for studying the molecular mechanisms involved in hESC fate determination. For these types of studies, feeder cell–dependent cultures of hESCs are often problematic because the physiology of the feeder cells is perturbed by the drug treatments or genetic modifications, which potentially obscures research outcomes. In this study, we evaluated three commonly used feeder-free culture conditions to determine whether they supported the undifferentiated growth of hESCs and to determine whether the hESCs grown in these conditions displayed gene expression patterns that were similar to the expression patterns of feeder cell-dependent hESCs. Our results demonstrate that hESCs grown in the three feeder-free conditions expressed undifferentiation marker genes as strongly as hESCs that were grown in the feeder- dependent cultures. Furthermore, genome-wide gene ex- pression profiles indicated that the gene expression patterns of hESCs that were grown under feeder-free or feeder- dependent culture conditions were highly similar. These results indicate that the feeder-free culture conditions support the undifferentiated growth of hESCs as effectively as the feeder-dependent culture conditions. Therefore, feeder-free culture conditions are potentially suitable for drug screening and for the genetic manipulation of hESCs in basic research. Keywords Feeder-free culture . Feeder-dependent culture . Human embryonic stem cells . Gene-expression profile Introduction Human embryonic stem cells (hESCs), established from the inner cell mass of blastocysts [1, 2], are able to indefinitely self-renew and differentiate into three germ layers [1–7]. Because of their pluripotent nature, hESCs are ideally suited for cell replacement therapies, drug development, and studies of early development. Although hESCs are typically grown on animal feeder cells, such as mouse embryonic fibroblasts (MEF) [1], human feeder cells, such as human fetal muscle and skin cells [8], human foreskin fibroblasts [9, 10], human uterine endometrial cells, and breast parenchymal cells [11], have also been successfully used for the undifferentiated growth of hESCs. In addition, a feeder-free culture of hESCs was reported for the first time in 2001 [12]. Feeder cell-free cultures of hESCs potentially provide several unique advantages for screening new drugs and for basic research. These advantages include minimal batch-to- batch variability, the high reproducibility of experimental outcomes, the easy standardization of experimental con- ditions, and the easy upscaling of the hESC cultures. Electronic supplementary material The online version of this article (doi:10.1007/s12015-010-9158-x) contains supplementary material, which is available to authorized users. T.-M. Yoon : B. Chang : H.-T. Kim : J.-H. Jee : D.-Y. Hwang (*) CHA Stem Cell Institute, Department of Biomedical Science, CHA University College of Medicine, Seoul 135-081, South Korea e-mail: hdy@cha.ac.kr D.-W. Kim Department of Physiology, Yonsei University College of Medicine, Seoul 120-752, South Korea Stem Cell Rev and Rep (2010) 6:425–437 DOI 10.1007/s12015-010-9158-x