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Derivation and maintenance of human embryonic stem cell line on human adult skin fibroblast feeder cells in serum replacement medium

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Abstract

Human embryonic stem (hES) cells were originally isolated and maintained on mouse embryonic fibroblast (MEF) feeder layers in the presence of fetal bovine serum (FBS). However, if the hES cells are to be used for therapeutic applications, it is preferable to regulatory authorities that they be derived and cultured in animal-free conditions to prevent mouse antigen contamination that would exacerbate an immune response to foreign proteins, and the potential risk of transmission of retroviral and other zoonotic pathogens to humans. As a step towards this goal, we derived a new hES cell line (MISCES-01) on human adult skin fibroblasts as feeder cells using serum replacement (SR) medium. The MISCES-01 cells have a normal diploid karyotype (46XX), express markers of pluripotency (OCT4, GCTM-2, TRA-1-60, TRA-1-81, SSEA-3, SSEA-4, and alkaline phosphatase) and following in vitro and in vivo differentiation, give rise to derivatives of the three primary germ layers. This cell line can be obtained for research purposes from the Australian Stem Cell Centre (http://www.stemcellcentre.edu.au).

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Acknowledgments

This research was funded by a grant to A. O. Trounson from the Australian Stem Cell Centre (ASCC), VIC, Australia. Human embryos were provided by Monash IVF Pty Ltd Clinic. We thank Ms. Bi Song (Monash University) for RT-PCR analysis, Dr. Mark Williamson and laboratory staff (Gribbles Pathology, Melbourne, Australia) for teratoma and pathology analyses, and Dr. Garey Dawson and laboratory staff (Monash Medical Centre, Southern Health, Australia) for cytogenetic analyses.

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Author notes

  1. Alan O. Trounson

    Present address: California Institute for Regenerative Medicine, 210 King Street, San Francisco, CA, 94107, USA

Authors and Affiliations

  1. Monash Immunology and Stem Cell Laboratories (MISCL), Monash Science Technology, Research and Innovation Precinct (STRIP), Monash University, Clayton, VIC, 3800, Australia

    Linh Nguyen, Karen Koh & Anna E. Michalska

  2. Research Office, Monash University, Clayton, VIC, 3800, Australia

    R. Tayfur Tecirlioglu

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  1. R. Tayfur Tecirlioglu
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  2. Linh Nguyen
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  3. Karen Koh
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  4. Alan O. Trounson
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  5. Anna E. Michalska
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Correspondence to Anna E. Michalska.

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Editor: J. Denry Sato

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Tecirlioglu, R.T., Nguyen, L., Koh, K. et al. Derivation and maintenance of human embryonic stem cell line on human adult skin fibroblast feeder cells in serum replacement medium. In Vitro Cell.Dev.Biol.-Animal 46, 231–235 (2010). https://doi.org/10.1007/s11626-010-9278-2

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  • DOI: https://doi.org/10.1007/s11626-010-9278-2

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