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    Engineered polymer-media interfaces for the long-term self-renewal of human embryonic stem cells

    Year: 2011

    Journal: Biomaterials, Volume 32, Issue 29, October 2011, Pages 6912-6919, 20110906

    Authors: Irwin E.F. 1, Gupta R. 1, Dashti D.C 1, Healy K.E. 1 2

    Last authors: Kevin E. Healy

    Organizations: 1 Department of Bioengineering, University of California at Berkeley, Berkeley, CA 94720, United States 2 Department of Material Science and Engineering, University of California at Berkeley, Berkeley, CA 94720, United States

    Country: USA, US, United States, United States of America, America

    We have developed a synthetic polymer interface for the long-term self-renewal of human embryonic stem cells (hESCs) in defined media. We successfully cultured hESCs on hydrogel interfaces of aminopropylmethacrylamide (APMAAm) for over 20 passages in chemically-defined mTeSR™1 media and demonstrated pluripotency of multiple hESC lines with immunostaining and quantitative RT-PCR studies. Results for hESC proliferation and pluripotency markers were both qualitatively and quantitatively similar to cells cultured on Matrigel™-coated substrates. Mechanistically, it was resolved that bovine serum albumin (BSA) in the mTeSR™1 media was critical for cell adhesion on APMAAm hydrogel interfaces. This study uniquely identified a robust long-term culture surface for the self-renewal of hESCs without the use of biologic coatings (e.g., peptides, proteins, or Matrigel™) in completely chemically-defined media that employed practical culturing techniques amenable to clinical-scale cell expansion.