caa a22 4500 445823070 CHVBK 20180317145250.0 cr unu---uuuuu 170323e20110501xx s 000 0 eng 10.1007/s10616-011-9344-y doi (NATIONALLICENCE)springer-10.1007/s10616-011-9344-y Sustained embryoid body formation and culture in a non-laborious three dimensional culture system for human embryonic stem cells [Elektronische Daten] [Johan Stenberg, Maria Elovsson, Raimund Strehl, Eva Kilmare, Johan Hyllner, Anders Lindahl] Pluripotent human embryonic stem cell (hESC) lines are a promising model system in developmental and tissue regeneration research. Differentiation of hESCs towards the three germ layers and finally tissue specific cell types is often performed through the formation of embryoid bodies (EBs) in suspension or hanging droplet culture systems. However, these systems are inefficient regarding embryoid body (EB) formation, structural support to the EB and long term differentiation capacity. The present study investigates if agarose, as a semi solid matrix, can facilitate EB formation and support differentiation of hESC lines. The results showed that agarose culture is able to enhance EB formation efficiency with 10% and increase EB growth by 300%. The agarose culture system was able to maintain expression of the three germ layers over 8weeks of culture. All of the four hESC lines tested developed EBs in the agarose system although with a histological heterogeneity between cell lines as well as within cell lines. In conclusion, a 3-D agarose culture of spherical hESC colonies improves EB formation and growth in a cost effective, stable and non-laborious technique. Springer Science+Business Media B.V., 2011 Human embryonic stem cell nationallicence In vitro differentiation nationallicence Embryoid body nationallicence Agarose nationallicence In vitro culture nationallicence hESC : Human embryonic stem cell nationallicence EB : Embryoid body nationallicence EBs : Embryoid bodies nationallicence SCID : Severe combined immunodeficiency nationallicence 3-D : Three dimensional nationallicence MEF : Mouse embryonic fibroblast nationallicence PBS : Phosphate buffered saline nationallicence AFP : Alpha fetoprotein nationallicence DAPI : 4′,6-diamidino-2-phenylindole nationallicence Bfgf : Basic fibroblast growth factor nationallicence PPD11 : p-Phenylenediamine anti-fade solution nationallicence IgG : Immuno globulin G nationallicence Stenberg Johan Department of Clinical Chemistry and Transfusion Medicine, Institute of Biomedicine, The Sahlgrenska Academy at University of Gothenburg, Sahlgrenska University Hospital, Klinisk Kemi, SU/Sahlgrenska, Bruna Stråket 16, 413 45, Gothenburg, Sweden aut Elovsson Maria Department of Clinical Chemistry and Transfusion Medicine, Institute of Biomedicine, The Sahlgrenska Academy at University of Gothenburg, Sahlgrenska University Hospital, Klinisk Kemi, SU/Sahlgrenska, Bruna Stråket 16, 413 45, Gothenburg, Sweden aut Strehl Raimund Cellartis AB, Arvid Wallgrens Backe 20, 413 46, Gothenburg, Sweden aut Kilmare Eva Cellartis AB, Arvid Wallgrens Backe 20, 413 46, Gothenburg, Sweden aut Hyllner Johan Cellartis AB, Arvid Wallgrens Backe 20, 413 46, Gothenburg, Sweden aut Lindahl Anders Department of Clinical Chemistry and Transfusion Medicine, Institute of Biomedicine, The Sahlgrenska Academy at University of Gothenburg, Sahlgrenska University Hospital, Klinisk Kemi, SU/Sahlgrenska, Bruna Stråket 16, 413 45, Gothenburg, Sweden aut Cytotechnology Springer Netherlands 63/3(2011-05-01), 227-237 0920-9069 63:3<227 2011 63 10616 https://doi.org/10.1007/s10616-011-9344-y text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1007/s10616-011-9344-y text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Stenberg Johan Department of Clinical Chemistry and Transfusion Medicine, Institute of Biomedicine, The Sahlgrenska Academy at University of Gothenburg, Sahlgrenska University Hospital, Klinisk Kemi, SU/Sahlgrenska, Bruna Stråket 16, 413 45, Gothenburg, Sweden aut NATIONALLICENCE 700 1- Elovsson Maria Department of Clinical Chemistry and Transfusion Medicine, Institute of Biomedicine, The Sahlgrenska Academy at University of Gothenburg, Sahlgrenska University Hospital, Klinisk Kemi, SU/Sahlgrenska, Bruna Stråket 16, 413 45, Gothenburg, Sweden aut NATIONALLICENCE 700 1- Strehl Raimund Cellartis AB, Arvid Wallgrens Backe 20, 413 46, Gothenburg, Sweden aut NATIONALLICENCE 700 1- Kilmare Eva Cellartis AB, Arvid Wallgrens Backe 20, 413 46, Gothenburg, Sweden aut NATIONALLICENCE 700 1- Hyllner Johan Cellartis AB, Arvid Wallgrens Backe 20, 413 46, Gothenburg, Sweden aut NATIONALLICENCE 700 1- Lindahl Anders Department of Clinical Chemistry and Transfusion Medicine, Institute of Biomedicine, The Sahlgrenska Academy at University of Gothenburg, Sahlgrenska University Hospital, Klinisk Kemi, SU/Sahlgrenska, Bruna Stråket 16, 413 45, Gothenburg, Sweden aut NATIONALLICENCE 773 0- Cytotechnology Springer Netherlands 63/3(2011-05-01), 227-237 0920-9069 63:3<227 2011 63 10616 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer