Polysulfone membranes coated with polymerized 3,4-dihydroxy-l-phenylalanine are a versatile and cost-effective synthetic substrate for defined long-term cultures of human pluripotent stem cells.
Clinical and industrial applications of human pluripotent stem cells (hPSC) require large amounts of cells that have been expanded under defined conditions. Labor-intensive techniques and ill-defined or expensive compounds and substrates are not applicable. Here we describe a chemically defined synthetic substrate consisting of polysulfone (PSF) membranes coated with polymerized 3,4-dihydroxy-l-phenylalanine (DOPA). DOPA/PSF is inexpensive and can be easily produced at various shapes and sizes. DOPA/PSF supports long-term self-renewal of undifferentiated human embryonic (hESC) and human induced pluripotent stem cells (hiPSC) under defined conditions. Pluripotency is maintained for at least 10 passages. Adhesion of hPSC to DOPA/PSF is mainly mediated by a specific integrin heterodimer. Proliferation and gene expression patterns on DOPA/PSF and control substrates are comparable. Labor-intensive cultivation methods and use of serum or coating with proteins are not required. Together, these features make DOPA/PSF attractive for applications where large-scale expansion of human pluripotent stem cells under defined conditions is essential.
Institute of Bioengineering and Nanotechnology , 31 Biopolis Way, The Nanos, Singapore 138669, Singapore.
Karthikeyan
K
Kandasamy
Karthikeyan
K
Narayanan
Ming
M
Ni
Chan
C
Du
Andrew C A
AC
Wan
Daniele
D
Zink
D018929:Cell Culture Techniques / Q000191:economics / Q000379:methods*; D049109:Cell Proliferation / Q000187:drug effects / Q000502:physiology; D003362:Cost-Benefit Analysis / Q000379:methods; D004295:Dihydroxyphenylalanine / Q000737:chemistry* / Q000191:economics / Q000494:pharmacology; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000187:drug effects* / Q000378:metabolism; D011108:Polymers / Q000737:chemistry* / Q000191:economics / Q000494:pharmacology; D013379:Substrate Specificity / Q000187:drug effects / Q000502:physiology; D013450:Sulfones / Q000737:chemistry* / Q000191:economics
D011108:Polymers; D013450:Sulfones; C017662:polysulfone P 1700; D004295:Dihydroxyphenylalanine