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

Nesrin Tüysüz, 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 R G Vries, Eric Braakman, Enrico Mastrobattista, Dirk-Jan Cornelissen, Hans Clevers, Derk Ten Berge

Onderzoeksoutput: Bijdrage aan wetenschappelijk tijdschrift/periodieke uitgaveArtikelWetenschappelijkpeer review

Samenvatting

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.

Originele taal-2Engels
Pagina's (van-tot)14578
TijdschriftNature Communications
Volume8
DOI's
StatusGepubliceerd - 06 mrt 2017

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