Feeder-independent culture of human embryonic stem cells

ABSTRACT We recently reported the development of TeSR1, a serum-free, animal product–free medium that supports the derivation and long-term feeder-independent culture of human embryonic stem

cells1. Although the derivation of new human embryonic stem cell lines in those defined conditions offered an important proof of principle, the costs of some of the defined components in

that culture system made it impractical for everyday research use. Here we describe modifications to the medium (mTeSR1) that include the use of animal-sourced proteins (bovine serum albumin

(BSA) and Matrigel) and cloned zebrafish basic fibroblast growth factor (zbFGF). We include a simple protocol that allows purification of up to 100 mg zbFGF in less than three days (Fig.

1), an amount sufficient to make 1,000 l of mTeSR1 medium. The modifications presented here make mTeSR1 practical for routine research use, and the protocols presented are those currently

used in our laboratory for standard human embryonic stem cell culture. *Note: In the version of this Protocol initially published, the references were numbered incorrectly. The error has

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institutional subscriptions * Read our FAQs * Contact customer support SIMILAR CONTENT BEING VIEWED BY OTHERS SIMPLIFICATION OF CULTURE CONDITIONS AND FEEDER-FREE EXPANSION OF BOVINE

EMBRYONIC STEM CELLS Article Open access 26 May 2021 TRUNCATED VITRONECTIN WITH E-CADHERIN ENABLES THE XENO-FREE DERIVATION OF HUMAN EMBRYONIC STEM CELLS Article Open access 12 September

2023 PRODUCTION AND CRYOPRESERVATION OF DEFINITIVE ENDODERM FROM HUMAN PLURIPOTENT STEM CELLS UNDER DEFINED AND SCALABLE CULTURE CONDITIONS Article 12 February 2021 CHANGE HISTORY * _ 24

AUGUST 2006 During production of the manuscript, the references were incorrectly numbered. The ordering within the text is fine and should remain the same, but the list of references needs

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described in this paper are those presently used by the Thomson Laboratory at the University of Wisconsin–Madison. We thank the many laboratory colleagues contributing to the preparation of

this manuscript, particularly J. Frane, D. Faupel and J. Antosiewicz for editorial review. This work was supported in part by US National Institutes of Health grants R24-RR017721 and

P20-GM069981, and funds from the University of Wisconsin Foundation. AUTHOR INFORMATION Author notes * Tenneille E Ludwig, Veit Bergendahl and Mark E Levenstein: These authors contributed

equally to the development of this protocol. AUTHORS AND AFFILIATIONS * Genome Center of Wisconsin, University of Wisconsin-Madison, 425 Henry Mall, Madison, 53706, Wisconsin, USA Tenneille

E Ludwig, Veit Bergendahl, Mitchell D Probasco & James A Thomson * WiCell Research Institute, P.O. Box 7365, Madison, 53707, Wisconsin, USA Tenneille E Ludwig, Mark E Levenstein & 

James A Thomson * National Primate Research Center, University of Wisconsin Graduate School, 1220 Capitol Court, Madison, 53705, Wisconsin, USA Tenneille E Ludwig, Junying Yu & James A

Thomson * Department of Anatomy, University of Wisconsin Medical School, 470 N. Charter Street, Madison, 53706, Wisconsin, USA James A Thomson Authors * Tenneille E Ludwig View author

publications You can also search for this author inPubMed Google Scholar * Veit Bergendahl View author publications You can also search for this author inPubMed Google Scholar * Mark E

Levenstein View author publications You can also search for this author inPubMed Google Scholar * Junying Yu View author publications You can also search for this author inPubMed Google

Scholar * Mitchell D Probasco View author publications You can also search for this author inPubMed Google Scholar * James A Thomson View author publications You can also search for this

author inPubMed Google Scholar CORRESPONDING AUTHOR Correspondence to Tenneille E Ludwig. ETHICS DECLARATIONS COMPETING INTERESTS J.A.T. owns stock in and has served on the Board of Cellular

Dynamics International (within the last two years), and owns stock in and serves on the Board of Stem Cell Products (within the last year). SUPPLEMENTARY INFORMATION SUPPLEMENTARY NOTE 1

(PDF 85 KB) RIGHTS AND PERMISSIONS Reprints and permissions ABOUT THIS ARTICLE CITE THIS ARTICLE Ludwig, T., Bergendahl, V., Levenstein, M. _et al._ Feeder-independent culture of human

embryonic stem cells. _Nat Methods_ 3, 637–646 (2006). https://doi.org/10.1038/nmeth902 Download citation * Published: 21 July 2006 * Issue Date: August 2006 * DOI:

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