Lipid-mediated Wnt protein stabilization enables serum-free culture of human organ stem cells

Nesrin Tuÿsüz, Louis Van Bloois, Stieneke Van Den Brink, Harry Begthel, Monique M.A. Verstegen, Luis J. Cruz, Lijian Hui, Luc J.W. Van Der Laan, Jeroen De Jonge, Robert Vries, Eric Braakman, Enrico Mastrobattista, Jan J. Cornelissen, Hans Clevers, Derk Ten Berge*

*Corresponding author for this work

Research output: Contribution to journalArticleAcademicpeer-review

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Wnt signalling proteins are essential for culture of human organ stem cells in organoids, but most Wnt protein formulations are poorly active in serum-free media. Here we show that purified Wnt3a protein is ineffective because it rapidly loses activity in culture media due to its hydrophobic nature, and its solubilization requires a detergent, CHAPS (3-[(3-cholamidopropyl) dimethylammonio]-1-propanesulfonate), that interferes with stem cell self-renewal. By stabilizing the Wnt3a protein using phospholipids and cholesterol as carriers, we address both problems: Wnt activity remains stable in serum-free media, while non-toxic carriers allow the use of high Wnt concentrations. Stabilized Wnt3a supports strongly increased self-renewal of organ and embryonic stem cells and the serum-free establishment of human organoids from healthy and diseased intestine and liver. Moreover, the lipophilicity of Wnt3a protein greatly facilitates its purification. Our findings remove a major obstacle impeding clinical applications of adult stem cells and offer advantages for all cell culture uses of Wnt3a protein.

Original languageEnglish
Article number14578
Number of pages11
JournalNature Communications
Publication statusPublished - 6 Mar 2017

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Publisher Copyright: © 2017 The Author(s).

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