caa a22 4500 445292008 CHVBK 20180317142531.0 cr unu---uuuuu 170323e20100401xx s 000 0 eng 10.1007/s11626-010-9297-z doi (NATIONALLICENCE)springer-10.1007/s11626-010-9297-z Comparison of defined culture systems for feeder cell free propagation of human embryonic stem cells [Elektronische Daten] [Veronika Akopian, Peter Andrews, Stephen Beil, Nissim Benvenisty, Jennifer Brehm, Megan Christie, Angela Ford, Victoria Fox, Paul Gokhale, Lyn Healy, Frida Holm, Outi Hovatta, Barbara Knowles, Tenneille Ludwig, Ronald McKay, Takamichi Miyazaki, Norio Nakatsuji, Steve Oh, Martin Pera, Janet Rossant, Glyn Stacey, Hirofumi Suemori] There are many reports of defined culture systems for the propagation of human embryonic stem cells in the absence of feeder cell support, but no previous study has undertaken a multi-laboratory comparison of these diverse methodologies. In this study, five separate laboratories, each with experience in human embryonic stem cell culture, used a panel of ten embryonic stem cell lines (including WA09 as an index cell line common to all laboratories) to assess eight cell culture methods, with propagation in the presence of Knockout Serum Replacer, FGF-2, and mouse embryonic fibroblast feeder cell layers serving as a positive control. The cultures were assessed for up to ten passages for attachment, death, and differentiated morphology by phase contrast microscopy, for growth by serial cell counts, and for maintenance of stem cell surface marker expression by flow cytometry. Of the eight culture systems, only the control and those based on two commercial media, mTeSR1 and STEMPRO, supported maintenance of most cell lines for ten passages. Cultures grown in the remaining media failed before this point due to lack of attachment, cell death, or overt cell differentiation. Possible explanations for relative success of the commercial formulations in this study, and the lack of success with other formulations from academic groups compared to previously published results, include: the complex combination of growth factors present in the commercial preparations; improved development, manufacture, and quality control in the commercial products; differences in epigenetic adaptation to culture in vitro between different ES cell lines grown in different laboratories. The Author(s), 2010 Human embryonic stem cell nationallicence Cell culture nationallicence Defined cell culture media nationallicence Comparative study nationallicence Akopian Veronika The Eli and Edythe Broad Center for Regenerative Medicine and Stem Cell Research at USC, Keck School of Medicine, University of Southern California, 90033, Los Angeles, CA, USA aut Andrews Peter Centre for Stem Cell Biology, Department of Biomedical Science, The University of Sheffield, S10 2TN, Sheffield, UK aut Beil Stephen The Eli and Edythe Broad Center for Regenerative Medicine and Stem Cell Research at USC, Keck School of Medicine, University of Southern California, 90033, Los Angeles, CA, USA aut Benvenisty Nissim Institute of Life Sciences, Department of Genetics, The Hebrew University of Jerusalem, Edmond J. Safra Campus, Givat Ram, 91904, Jerusalem, Israel aut Brehm Jennifer WiCell Research Institute, P.O. Box7365, 53707-7365, Madison, WI, USA aut Christie Megan UK Stem Cell Bank, Division of Cell Biology and Imaging, National Institute for Biological Standards and Control-Health Protection Agency, South Mimms, EN6 3QG, Herts, UK aut Ford Angela Centre for Stem Cell Biology, Department of Biomedical Science, The University of Sheffield, S10 2TN, Sheffield, UK aut Fox Victoria The Eli and Edythe Broad Center for Regenerative Medicine and Stem Cell Research at USC, Keck School of Medicine, University of Southern California, 90033, Los Angeles, CA, USA aut Gokhale Paul Centre for Stem Cell Biology, Department of Biomedical Science, The University of Sheffield, S10 2TN, Sheffield, UK aut Healy Lyn UK Stem Cell Bank, Division of Cell Biology and Imaging, National Institute for Biological Standards and Control-Health Protection Agency, South Mimms, EN6 3QG, Herts, UK aut Holm Frida Karolinska Institutet, Department of Clinical Science, Technology and Intervention, Karolinska University Hospital, SE 141 86, Stockholm, Sweden aut Hovatta Outi Karolinska Institutet, Department of Clinical Science, Technology and Intervention, Karolinska University Hospital, SE 141 86, Stockholm, Sweden aut Knowles Barbara Institute of Medical Biology 8A Biomedical Grove, #06-06 Immunos, 138648, Singapore, Republic of Singapore aut Ludwig Tenneille WiCell Research Institute, P.O. Box7365, 53707-7365, Madison, WI, USA aut McKay Ronald NIH Stem Cell Unit, National Institutes of Health, Bldg 35/2B-213, MSC 3703, 9000 Rockville Pike, 20892, Bethesda, MD, USA aut Miyazaki Takamichi Institute for Frontier Medical Sciences, Kyoto University, 606-8507, Kyoto, Japan aut Nakatsuji Norio Institute for Frontier Medical Sciences, Kyoto University, 606-8507, Kyoto, Japan aut Oh Steve Bioprocessing Technology Institute, Agency for Science Technology and Research (A*STAR), 20 Biopolis Way, #06-01 Centros, 138668, Singapore, Republic of Singapore aut Pera Martin The Eli and Edythe Broad Center for Regenerative Medicine and Stem Cell Research at USC, Keck School of Medicine, University of Southern California, 90033, Los Angeles, CA, USA aut Rossant Janet Developmental Biology Program, The Hospital for Sick Children, TMDT Building, Room 13-305, 101 College Street, M5G 1L7, Toronto, ON, Canada aut Stacey Glyn UK Stem Cell Bank, Division of Cell Biology and Imaging, National Institute for Biological Standards and Control-Health Protection Agency, South Mimms, EN6 3QG, Herts, UK aut Suemori Hirofumi Institute for Frontier Medical Sciences, Kyoto University, 606-8507, Kyoto, Japan aut In Vitro Cellular & Developmental Biology - Animal Springer-Verlag 46/3-4(2010-04-01), 247-258 1071-2690 46:3-4<247 2010 46 11626 https://doi.org/10.1007/s11626-010-9297-z text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1007/s11626-010-9297-z text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Akopian Veronika The Eli and Edythe Broad Center for Regenerative Medicine and Stem Cell Research at USC, Keck School of Medicine, University of Southern California, 90033, Los Angeles, CA, USA aut NATIONALLICENCE 700 1- Andrews Peter Centre for Stem Cell Biology, Department of Biomedical Science, The University of Sheffield, S10 2TN, Sheffield, UK aut NATIONALLICENCE 700 1- Beil Stephen The Eli and Edythe Broad Center for Regenerative Medicine and Stem Cell Research at USC, Keck School of Medicine, University of Southern California, 90033, Los Angeles, CA, USA aut NATIONALLICENCE 700 1- Benvenisty Nissim Institute of Life Sciences, Department of Genetics, The Hebrew University of Jerusalem, Edmond J. Safra Campus, Givat Ram, 91904, Jerusalem, Israel aut NATIONALLICENCE 700 1- Brehm Jennifer WiCell Research Institute, P.O. Box7365, 53707-7365, Madison, WI, USA aut NATIONALLICENCE 700 1- Christie Megan UK Stem Cell Bank, Division of Cell Biology and Imaging, National Institute for Biological Standards and Control-Health Protection Agency, South Mimms, EN6 3QG, Herts, UK aut NATIONALLICENCE 700 1- Ford Angela Centre for Stem Cell Biology, Department of Biomedical Science, The University of Sheffield, S10 2TN, Sheffield, UK aut NATIONALLICENCE 700 1- Fox Victoria The Eli and Edythe Broad Center for Regenerative Medicine and Stem Cell Research at USC, Keck School of Medicine, University of Southern California, 90033, Los Angeles, CA, USA aut NATIONALLICENCE 700 1- Gokhale Paul Centre for Stem Cell Biology, Department of Biomedical Science, The University of Sheffield, S10 2TN, Sheffield, UK aut NATIONALLICENCE 700 1- Healy Lyn UK Stem Cell Bank, Division of Cell Biology and Imaging, National Institute for Biological Standards and Control-Health Protection Agency, South Mimms, EN6 3QG, Herts, UK aut NATIONALLICENCE 700 1- Holm Frida Karolinska Institutet, Department of Clinical Science, Technology and Intervention, Karolinska University Hospital, SE 141 86, Stockholm, Sweden aut NATIONALLICENCE 700 1- Hovatta Outi Karolinska Institutet, Department of Clinical Science, Technology and Intervention, Karolinska University Hospital, SE 141 86, Stockholm, Sweden aut NATIONALLICENCE 700 1- Knowles Barbara Institute of Medical Biology 8A Biomedical Grove, #06-06 Immunos, 138648, Singapore, Republic of Singapore aut NATIONALLICENCE 700 1- Ludwig Tenneille WiCell Research Institute, P.O. Box7365, 53707-7365, Madison, WI, USA aut NATIONALLICENCE 700 1- McKay Ronald NIH Stem Cell Unit, National Institutes of Health, Bldg 35/2B-213, MSC 3703, 9000 Rockville Pike, 20892, Bethesda, MD, USA aut NATIONALLICENCE 700 1- Miyazaki Takamichi Institute for Frontier Medical Sciences, Kyoto University, 606-8507, Kyoto, Japan aut NATIONALLICENCE 700 1- Nakatsuji Norio Institute for Frontier Medical Sciences, Kyoto University, 606-8507, Kyoto, Japan aut NATIONALLICENCE 700 1- Oh Steve Bioprocessing Technology Institute, Agency for Science Technology and Research (A*STAR), 20 Biopolis Way, #06-01 Centros, 138668, Singapore, Republic of Singapore aut NATIONALLICENCE 700 1- Pera Martin The Eli and Edythe Broad Center for Regenerative Medicine and Stem Cell Research at USC, Keck School of Medicine, University of Southern California, 90033, Los Angeles, CA, USA aut NATIONALLICENCE 700 1- Rossant Janet Developmental Biology Program, The Hospital for Sick Children, TMDT Building, Room 13-305, 101 College Street, M5G 1L7, Toronto, ON, Canada aut NATIONALLICENCE 700 1- Stacey Glyn UK Stem Cell Bank, Division of Cell Biology and Imaging, National Institute for Biological Standards and Control-Health Protection Agency, South Mimms, EN6 3QG, Herts, UK aut NATIONALLICENCE 700 1- Suemori Hirofumi Institute for Frontier Medical Sciences, Kyoto University, 606-8507, Kyoto, Japan aut NATIONALLICENCE 773 0- In Vitro Cellular & Developmental Biology - Animal Springer-Verlag 46/3-4(2010-04-01), 247-258 1071-2690 46:3-4<247 2010 46 11626 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer