title: Characterization of Phenotypic and Transcriptional Differences in Human Pluripotent Stem Cells under 2D and 3D Culture Conditions.
abstract: Human pluripotent stem cells hold great promise for applications in drug discovery and regenerative medicine. Microfluidic technology is a promising approach for creating artificial microenvironments; however, although a proper 3D microenvironment is required to achieve robust control of cellular phenotypes, most current microfluidic devices provide only 2D cell culture and do not allow tuning of physical and chemical environmental cues simultaneously. Here, the authors report a 3D cellular microenvironment plate (3D-CEP), which consists of a microfluidic device filled with thermoresponsive poly(N-isopropylacrylamide)-β-poly(ethylene glycol) hydrogel (HG), which enables systematic tuning of both chemical and physical environmental cues as well as in situ cell monitoring. The authors show that H9 human embryonic stem cells (hESCs) and 253G1 human induced pluripotent stem cells in the HG/3D-CEP system maintain their pluripotent marker expression under HG/3D-CEP self-renewing conditions. Additionally, global gene expression analyses are used to elucidate small variations among different test environments. Interestingly, the authors find that treatment of H9 hESCs under HG/3D-CEP self-renewing conditions results in initiation of entry into the neural differentiation process by induction of PAX3 and OTX1 expression. The authors believe that this HG/3D-CEP system will serve as a versatile platform for developing targeted functional cell lines and facilitate advances in drug screening and regenerative medicine.
authors: Institute for Integrated Cell-Material Sciences (WPI-iCeMS), Kyoto University, Kyoto, 6068501, Japan.
authors: Ken-Ichiro
authors: KI
authors: Kamei
authors: Institute for Integrated Cell-Material Sciences (WPI-iCeMS), Kyoto University, Kyoto, 6068501, Japan.
authors: Yoshie
authors: Y
authors: Koyama
authors: Institute for Integrated Cell-Material Sciences (WPI-iCeMS), Kyoto University, Kyoto, 6068501, Japan.
authors: Yumie
authors: Y
authors: Tokunaga
authors: Institute for Integrated Cell-Material Sciences (WPI-iCeMS), Kyoto University, Kyoto, 6068501, Japan.
authors: Yasumasa
authors: Y
authors: Mashimo
authors: Institute for Integrated Cell-Material Sciences (WPI-iCeMS), Kyoto University, Kyoto, 6068501, Japan.
authors: Momoko
authors: M
authors: Yoshioka
authors: Institute for Integrated Cell-Material Sciences (WPI-iCeMS), Kyoto University, Kyoto, 6068501, Japan.
authors: Christopher
authors: C
authors: Fockenberg
authors: Australia Institute for Biotechnology and Nanotechnology (AIBN), University of Queensland, Brisbane, QLD, 4072, Australia.|Department of Anatomy and Neuroscience, University of Melbourne, Melbourne, Vic, 3010, Australia.
authors: Rowland
authors: R
authors: Mosbergen
authors: Australia Institute for Biotechnology and Nanotechnology (AIBN), University of Queensland, Brisbane, QLD, 4072, Australia.
authors: Othmar
authors: O
authors: Korn
authors: Australia Institute for Biotechnology and Nanotechnology (AIBN), University of Queensland, Brisbane, QLD, 4072, Australia.|Department of Anatomy and Neuroscience, University of Melbourne, Melbourne, Vic, 3010, Australia.
authors: Christine
authors: C
authors: Wells
authors: Institute for Integrated Cell-Material Sciences (WPI-iCeMS), Kyoto University, Kyoto, 6068501, Japan.|Ecole Normale Supérieure, CNRS-ENS-UPMC UMR 8640, 24 Rue L'homond, Paris, 75005, France.
authors: Yong
authors: Y
authors: Chen
mesh_terms: D000180:Acrylic Resins / Q000008:administration & dosage / Q000737:chemistry; D015415:Biomarkers / Q000378:metabolism; D018929:Cell Culture Techniques / Q000379:methods; D002454:Cell Differentiation / Q000502:physiology; D002478:Cells, Cultured; D060833:Cellular Microenvironment / Q000502:physiology; D000066449:Human Embryonic Stem Cells / Q000166:cytology; D006801:Humans; D020136:Hydrogel, Polyethylene Glycol Dimethacrylate / Q000008:administration & dosage / Q000737:chemistry; D056656:Lab-On-A-Chip Devices; D039904:Pluripotent Stem Cells / Q000166:cytology* / Q000378:metabolism; D011092:Polyethylene Glycols / Q000008:administration & dosage / Q000737:chemistry; D044968:Regenerative Medicine / Q000379:methods; D014158:Transcription, Genetic / Q000502:physiology
chemical_list: D000180:Acrylic Resins; D015415:Biomarkers; C052970:poly-N-isopropylacrylamide; D020136:Hydrogel, Polyethylene Glycol Dimethacrylate; D011092:Polyethylene Glycols
keywords: 3D cellular microenvironment; human pluripotent stem cells; hydrogels; microfluidic devices; self-renewal