Human Embryonic Stem Cell Protocols pp 61-69 | Cite as
Efficient Expansion of Dissociated Human Pluripotent Stem Cells Using a Synthetic Substrate
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Abstract
Human pluripotent stem cells (hPSCs), including human embryonic stem cells and human-induced pluripotent stem cells, are a renewable cell source for a wide range of applications in regenerative medicine and useful tools for human disease modeling and drug discovery. For these purposes, large numbers of high-quality cells are essential. Recently, we showed that a biological substrate, recombinant E8 fragments of laminin isoforms, sustains long-term self-renewal of hPSCs in defined, xeno-free medium with dissociated single-cell passaging. Here, we describe a modified culture system with similar performance to efficiently expand hPSCs under defined, xeno-free conditions using a non-biological synthetic substrate.
Keywords:
Human embryonic stem cells Synthetic substrate Defined culture Xeno-free culture Culture substrateNotes
Acknowledgments
I thank Ms. Fumi Kashigi and Mari Hamao for their valuable technical assistance and the members of the laboratories of N. Nakatsuji for insightful discussions. This work was supported by a Grant-in-Aid for Scientific Research and partially supported by the New Energy and Industrial Technology Development Organization (NEDO) of Japan.
References
- 1.Thomson JA, Itskovitz-Eldor J, Shapiro SS, Waknitz MA, Swiergiel JJ, Marshall VS, Jones JM (1998) Embryonic stem cell lines derived from human blastocysts. Science 282:1145–1147PubMedCrossRefGoogle Scholar
- 2.Takahashi K, Tanabe K, Ohnuki M, Narita M, Ichisaka T, Tomoda K, Yamanaka S (2007) Induction of pluripotent stem cells from adult human fibroblasts by defined factors. Cell 131:861–872PubMedCrossRefGoogle Scholar
- 3.Yu J, Vodyanik MA, Smuga-Otto K, Antosiewicz-Bourget J, Frane JL, Tian S, Nie J, Jonsdottir GA, Ruotti V, Stewart R, Slukvin II, Thomson JA (2007) Induced pluripotent stem cell lines derived from human somatic cells. Science 318:1917–1920PubMedCrossRefGoogle Scholar
- 4.Segers VF, Lee RT (2008) Stem-cell therapy for cardiac disease. Nature 451:937–942PubMedCrossRefGoogle Scholar
- 5.Serra M, Brito C, Correia C, Alves PM (2012) Process engineering of human pluripotent stem cells for clinical application. Trends Biotechnol 30:350–359PubMedCrossRefGoogle Scholar
- 6.Soldner F, Laganiere J, Cheng AW et al (2011) Generation of isogenic pluripotent stem cells differing exclusively at two early onset Parkinson point mutations. Cell 146:318–331PubMedCentralPubMedCrossRefGoogle Scholar
- 7.Amit M, Carpenter MK, Inokuma MS, Chiu CP, Harris CP, Waknitz MA, Itskovitz-Eldor J, Thomson JA (2000) Clonally derived human embryonic stem cell lines maintain pluripotency and proliferative potential for prolonged periods of culture. Dev Biol 227:271–278PubMedCrossRefGoogle Scholar
- 8.Pyle AD, Lock LF, Donovan PJ (2006) Neurotrophins mediate human embryonic stem cell survival. Nat Biotechnol 24:344–350PubMedCrossRefGoogle Scholar
- 9.Miyazaki T, Futaki S, Suemori H, Taniguchi Y, Yamada M, Kawasaki M, Hayashi M, Kumagai H, Nakatsuji N, Sekiguchi K, Kawase E (2012) Laminin E8 fragments support efficient adhesion and expansion of dissociated human pluripotent stem cells. Nat Commun 3:1236PubMedCentralPubMedCrossRefGoogle Scholar
- 10.Melkoumian Z, Weber JL, Weber DM et al (2010) Synthetic peptide-acrylate surfaces for long-term self-renewal and cardiomyocyte differentiation of human embryonic stem cells. Nat Biotechnol 28(606–610):606–610PubMedCrossRefGoogle Scholar