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

doi:10.1038/ncomms14578

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

Hubrecht Institute

Research output: Contribution to journal/periodical › Article › Scientific › peer-review

57 Citations (Scopus)

Abstract

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 language	English
Pages (from-to)	14578
Journal	Nature Communications
Volume	8
DOIs	https://doi.org/10.1038/ncomms14578
Publication status	Published - 06 Mar 2017

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Journal Article

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10.1038/ncomms14578

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Tüysüz, N., van Bloois, L., van den Brink, S., Begthel, H., Verstegen, M. M. A., Cruz, L. J., Hui, L., van der Laan, L. J. W., de Jonge, J., Vries, R. R. G., Braakman, E., Mastrobattista, E., Cornelissen, D.-J., Clevers, H., & Ten Berge, D. (2017). Lipid-mediated Wnt protein stabilization enables serum-free culture of human organ stem cells. Nature Communications, 8, 14578. https://doi.org/10.1038/ncomms14578

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Tüysüz, N, van Bloois, L, van den Brink, S, Begthel, H, Verstegen, MMA, Cruz, LJ, Hui, L, van der Laan, LJW, de Jonge, J, Vries, RRG, Braakman, E, Mastrobattista, E, Cornelissen, D-J, Clevers, H & Ten Berge, D 2017, 'Lipid-mediated Wnt protein stabilization enables serum-free culture of human organ stem cells', Nature Communications, vol. 8, pp. 14578. https://doi.org/10.1038/ncomms14578

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AU - Tüysüz, Nesrin

AU - van Bloois, Louis

AU - van den Brink, Stieneke

AU - Begthel, Harry

AU - Verstegen, Monique M A

AU - Cruz, Luis J

AU - Hui, Lijian

AU - van der Laan, Luc J W

AU - de Jonge, Jeroen

AU - Vries, Robert R G

AU - Braakman, Eric

AU - Mastrobattista, Enrico

AU - Cornelissen, Dirk-Jan

AU - Clevers, Hans

AU - Ten Berge, Derk

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N2 - 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.

AB - 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.

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DO - 10.1038/ncomms14578

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SN - 2041-1723

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JO - Nature Communications

JF - Nature Communications

ER -

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

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