StemBond hydrogels control the mechanical microenvironment for pluripotent stem cells

Celine Labouesse; Bao Xiu Tan; Chibeza C. Agley; Moritz Hofer; Alexander K. Winkel; Giuliano G. Stirparo; Hannah T. Stuart; Christophe M. Verstreken; Carla Mulas; William Mansfield; Paul Bertone; Kristian Franze; Jose C. R. Silva; Kevin J. Chalut

doi:10.1038/s41467-021-26236-5

StemBond hydrogels control the mechanical microenvironment for pluripotent stem cells

Celine Labouesse, Bao Xiu Tan, Chibeza C. Agley, Moritz Hofer, Alexander K. Winkel, Giuliano G. Stirparo, Hannah T. Stuart, Christophe M. Verstreken, Carla Mulas, William Mansfield, Paul Bertone, Kristian Franze, Jose C. R. Silva, Kevin J. Chalut

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25 Citations (Scopus)

Abstract

Studies of mechanical signalling are typically performed by comparing cells cultured on soft and stiff hydrogel-based substrates. However, it is challenging to independently and robustly control both substrate stiffness and extracellular matrix tethering to substrates, making matrix tethering a potentially confounding variable in mechanical signalling investigations. Moreover, unstable matrix tethering can lead to poor cell attachment and weak engagement of cell adhesions. To address this, we developed StemBond hydrogels, a hydrogel in which matrix tethering is robust and can be varied independently of stiffness. We validate StemBond hydrogels by showing that they provide an optimal system for culturing mouse and human pluripotent stem cells. We further show how soft StemBond hydrogels modulate stem cell function, partly through stiffness-sensitive ERK signalling. Our findings underline how substrate mechanics impact mechanosensitive signalling pathways regulating self-renewal and differentiation, indicating that optimising the complete mechanical microenvironment will offer greater control over stem cell fate specification.

Original language	English
Article number	6132
Journal	Nature Communications
Volume	12
Issue number	1
DOIs	https://doi.org/10.1038/s41467-021-26236-5
Publication status	Published - 21 Oct 2021

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Labouesse, C., Tan, B. X., Agley, C. C., Hofer, M., Winkel, A. K., Stirparo, G. G., Stuart, H. T., Verstreken, C. M., Mulas, C., Mansfield, W., Bertone, P., Franze, K., Silva, J. C. R., & Chalut, K. J. (2021). StemBond hydrogels control the mechanical microenvironment for pluripotent stem cells. Nature Communications, 12(1), Article 6132. https://doi.org/10.1038/s41467-021-26236-5

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title = "StemBond hydrogels control the mechanical microenvironment for pluripotent stem cells",

abstract = "Studies of mechanical signalling are typically performed by comparing cells cultured on soft and stiff hydrogel-based substrates. However, it is challenging to independently and robustly control both substrate stiffness and extracellular matrix tethering to substrates, making matrix tethering a potentially confounding variable in mechanical signalling investigations. Moreover, unstable matrix tethering can lead to poor cell attachment and weak engagement of cell adhesions. To address this, we developed StemBond hydrogels, a hydrogel in which matrix tethering is robust and can be varied independently of stiffness. We validate StemBond hydrogels by showing that they provide an optimal system for culturing mouse and human pluripotent stem cells. We further show how soft StemBond hydrogels modulate stem cell function, partly through stiffness-sensitive ERK signalling. Our findings underline how substrate mechanics impact mechanosensitive signalling pathways regulating self-renewal and differentiation, indicating that optimising the complete mechanical microenvironment will offer greater control over stem cell fate specification.",

author = "Celine Labouesse and Tan, {Bao Xiu} and Agley, {Chibeza C.} and Moritz Hofer and Winkel, {Alexander K.} and Stirparo, {Giuliano G.} and Stuart, {Hannah T.} and Verstreken, {Christophe M.} and Carla Mulas and William Mansfield and Paul Bertone and Kristian Franze and Silva, {Jose C. R.} and Chalut, {Kevin J.}",

note = "Funding Information: We are grateful to Sarah Pallett for excellent support in all experiments, Peter Humphreys for assistance with imaging and imaging analysis, Sally Lees and staff for support in tissue culture, Maike Paramor and staff for preparation of sequencing libraries, Michael A. Barber for alignment of sequencing data, G. Chu and staff for animal husbandry, Ivan B. Dimov and Amelia Joy Thompson for help with AFM measurements. Rosa26-CreERT2+/+ cells were a kind gift from the lab of Bon-Kyoung Koo, mESCs and hPSC cell lines were a kind gift from the Smith lab. Illumina RNA sequencing was performed at the CIGC, Cambridge. This work was financially supported by core funding from the UKRI Medical Research Council [MC_PC_17230] and the Wellcome Trust [203151/Z/16/Z] to the Wellcome-MRC Cambridge Stem Cell Institute, and individual grants to K.J.C. and J.C.R.S. from the Medical Research Council (MR/M011089/1) and to K.J.C. from the European Research Council (772798). B.X.T. is supported by a Wellcome 4-year Ph.D. grant. J.C.R.S. is a Wellcome Trust Senior Research Fellow (WT101861). K.J.C. is a Royal Society University Research Fellow. K.F. received support from MRC Career Development Award (G1100312/1) and ERC (772426) and Alexander von Humboldt Foundation. For the purpose of open access, the author has applied a CC BY public copyright licence to any Author Accepted Manuscript version arising from this submission. Publisher Copyright: {\textcopyright} 2021, The Author(s).",

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doi = "10.1038/s41467-021-26236-5",

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AU - Labouesse, Celine

AU - Tan, Bao Xiu

AU - Agley, Chibeza C.

AU - Hofer, Moritz

AU - Winkel, Alexander K.

AU - Stirparo, Giuliano G.

AU - Stuart, Hannah T.

AU - Verstreken, Christophe M.

AU - Mulas, Carla

AU - Mansfield, William

AU - Bertone, Paul

AU - Franze, Kristian

AU - Silva, Jose C. R.

AU - Chalut, Kevin J.

N1 - Funding Information: We are grateful to Sarah Pallett for excellent support in all experiments, Peter Humphreys for assistance with imaging and imaging analysis, Sally Lees and staff for support in tissue culture, Maike Paramor and staff for preparation of sequencing libraries, Michael A. Barber for alignment of sequencing data, G. Chu and staff for animal husbandry, Ivan B. Dimov and Amelia Joy Thompson for help with AFM measurements. Rosa26-CreERT2+/+ cells were a kind gift from the lab of Bon-Kyoung Koo, mESCs and hPSC cell lines were a kind gift from the Smith lab. Illumina RNA sequencing was performed at the CIGC, Cambridge. This work was financially supported by core funding from the UKRI Medical Research Council [MC_PC_17230] and the Wellcome Trust [203151/Z/16/Z] to the Wellcome-MRC Cambridge Stem Cell Institute, and individual grants to K.J.C. and J.C.R.S. from the Medical Research Council (MR/M011089/1) and to K.J.C. from the European Research Council (772798). B.X.T. is supported by a Wellcome 4-year Ph.D. grant. J.C.R.S. is a Wellcome Trust Senior Research Fellow (WT101861). K.J.C. is a Royal Society University Research Fellow. K.F. received support from MRC Career Development Award (G1100312/1) and ERC (772426) and Alexander von Humboldt Foundation. For the purpose of open access, the author has applied a CC BY public copyright licence to any Author Accepted Manuscript version arising from this submission. Publisher Copyright: © 2021, The Author(s).

PY - 2021/10/21

Y1 - 2021/10/21

N2 - Studies of mechanical signalling are typically performed by comparing cells cultured on soft and stiff hydrogel-based substrates. However, it is challenging to independently and robustly control both substrate stiffness and extracellular matrix tethering to substrates, making matrix tethering a potentially confounding variable in mechanical signalling investigations. Moreover, unstable matrix tethering can lead to poor cell attachment and weak engagement of cell adhesions. To address this, we developed StemBond hydrogels, a hydrogel in which matrix tethering is robust and can be varied independently of stiffness. We validate StemBond hydrogels by showing that they provide an optimal system for culturing mouse and human pluripotent stem cells. We further show how soft StemBond hydrogels modulate stem cell function, partly through stiffness-sensitive ERK signalling. Our findings underline how substrate mechanics impact mechanosensitive signalling pathways regulating self-renewal and differentiation, indicating that optimising the complete mechanical microenvironment will offer greater control over stem cell fate specification.

AB - Studies of mechanical signalling are typically performed by comparing cells cultured on soft and stiff hydrogel-based substrates. However, it is challenging to independently and robustly control both substrate stiffness and extracellular matrix tethering to substrates, making matrix tethering a potentially confounding variable in mechanical signalling investigations. Moreover, unstable matrix tethering can lead to poor cell attachment and weak engagement of cell adhesions. To address this, we developed StemBond hydrogels, a hydrogel in which matrix tethering is robust and can be varied independently of stiffness. We validate StemBond hydrogels by showing that they provide an optimal system for culturing mouse and human pluripotent stem cells. We further show how soft StemBond hydrogels modulate stem cell function, partly through stiffness-sensitive ERK signalling. Our findings underline how substrate mechanics impact mechanosensitive signalling pathways regulating self-renewal and differentiation, indicating that optimising the complete mechanical microenvironment will offer greater control over stem cell fate specification.

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StemBond hydrogels control the mechanical microenvironment for pluripotent stem cells

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