We describe a system for culturing human embryonic stem (hES) cells and induced pluripotent stem (iPS) cells on a recombinant form of human laminin-511, a component of the natural hES cell niche. The system is devoid of animal products and feeder cells and contains only one undefined component, human albumin. The hES cells self-renewed with normal karyotype for at least 4 months (20 passages), after which the cells could produce teratomas containing cell lineages of all three germ layers. When plated on laminin-511 in small clumps, hES cells spread out in a monolayer, maintaining cellular homogeneity with approximately 97% OCT4-positive cells. Adhesion of hES cells was dependent on α6β1 integrin. The use of homogeneous monolayer hES or iPS cell cultures provides more controllable conditions for the design of differentiation methods. This xeno-free and feeder-free system may be useful for the development of cell lineages for therapeutic purposes.
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We thank A.-S. Nilsson and A.-M. Strömberg for excellent technical assistance, C. Cowan for collaboration in procuring the iPS cell lines, F. Holm and R. Bergström for their help with the cell cultures and D. Baker for carrying out karyotyping of hES cells. This work was supported in part by grants from the Knut and Alice Wallenberg Foundation (K.T.), the Novo Nordisk Foundation (K.T.), the Söderberg's Foundation (K.T.), the Swedish Research Council (K.T., O.H.), the Swedish Cancer Foundation (K.T.), the Harvard Stem Cell Institute (K.R.C.) and the Leducq Foundation (K.R.C.). E.M.H. is a Wenner-Gren Foundation fellow. EU: ESTOOLS (O.H.) has not been used for the derivation of new hES cell lines.
K.T. and S.R. are shareholders in BioLamina.
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Rodin, S., Domogatskaya, A., Ström, S. et al. Long-term self-renewal of human pluripotent stem cells on human recombinant laminin-511. Nat Biotechnol 28, 611–615 (2010) doi:10.1038/nbt.1620
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