Volume adaptation controls stem cell mechanotransduction

Luke Major, Andrew W Holle, Jennifer L Young, Matt Hepburn, Kwanghee Jeong, Ian Chin, Rowan Sanderson, Ji Hoon Jeong, Zach Aman, Brendan Kennedy, Yongsung Hwang, Dong-Wook Han, Hyun Woo Park, Kun-Liang Guan, Joachim P Spatz, Yu Suk Choi

Research output: Contribution to journalArticle

2 Citations (Scopus)

Abstract

Recent studies have found discordant mechanosensitive outcomes when comparing 2D and 3D, highlighting the need for tools to study mechanotransduction in 3D across a wide spectrum of stiffness. A gelatin methacryloyl (GelMA) hydrogel with a continuous stiffness gradient ranging from 5 to 38 kPa was developed to recapitulate physiological stiffness conditions. Adipose-derived stem cells (ASCs) were encapsulated in this hydrogel and their morphological characteristics and expression of both mechanosensitive proteins (Lamin A, YAP, and MRTFa) and differentiation markers (PPARγ and RUNX2) were analyzed. Low stiffness regions (~8 kPa) permitted increased cellular and nuclear volume and enhanced mechanosensitive protein localization in the nucleus. This trend was reversed in high stiffness regions (~30 kPa), where decreased cellular and nuclear volumes and reduced mechanosensitive protein nuclear localization were observed. Interestingly, cells in soft regions exhibited enhanced osteogenic RUNX2 expression, while those in stiff regions upregulated the adipogenic regulator PPARγ, suggesting that volume, not substrate stiffness, is sufficient to drive 3D stem cell differentiation. Inhibition of myosin II (Blebbistatin) and ROCK (Y-27632), both key drivers of actomyosin contractility, resulted in reduced cell volume, especially in low stiffness regions, causing a decorrelation between volume expansion and mechanosensitive protein localization. Constitutively active and inactive forms of the canonical downstream mechanotransduction effector TAZ were stably transfected into ASCs. Activated TAZ resulted in higher cellular volume despite increasing stiffness and a consistent, stiffness-independent translocation of YAP and MRTFa into the nucleus. Thus, volume adaptation as a function of 3D matrix stiffness can control stem cell mechanotransduction and differentiation.
Original languageEnglish
Pages (from-to)45520-45530
Number of pages11
JournalACS Applied Materials and Interfaces
Volume11
Issue number49
Early online date12 Nov 2019
DOIs
Publication statusPublished - 11 Dec 2019

Fingerprint

Stem cells
Stiffness
Peroxisome Proliferator-Activated Receptors
Hydrogel
Proteins
Hydrogels
Lamin Type A
Myosin Type II
Actomyosin
Differentiation Antigens
Stiffness matrix
Gelatin
Nuclear Proteins
Substrates

Cite this

Major, Luke ; Holle, Andrew W ; Young, Jennifer L ; Hepburn, Matt ; Jeong, Kwanghee ; Chin, Ian ; Sanderson, Rowan ; Jeong, Ji Hoon ; Aman, Zach ; Kennedy, Brendan ; Hwang, Yongsung ; Han, Dong-Wook ; Park, Hyun Woo ; Guan, Kun-Liang ; Spatz, Joachim P ; Choi, Yu Suk. / Volume adaptation controls stem cell mechanotransduction. In: ACS Applied Materials and Interfaces. 2019 ; Vol. 11, No. 49. pp. 45520-45530.
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title = "Volume adaptation controls stem cell mechanotransduction",
abstract = "Recent studies have found discordant mechanosensitive outcomes when comparing 2D and 3D, highlighting the need for tools to study mechanotransduction in 3D across a wide spectrum of stiffness. A gelatin methacryloyl (GelMA) hydrogel with a continuous stiffness gradient ranging from 5 to 38 kPa was developed to recapitulate physiological stiffness conditions. Adipose-derived stem cells (ASCs) were encapsulated in this hydrogel and their morphological characteristics and expression of both mechanosensitive proteins (Lamin A, YAP, and MRTFa) and differentiation markers (PPARγ and RUNX2) were analyzed. Low stiffness regions (~8 kPa) permitted increased cellular and nuclear volume and enhanced mechanosensitive protein localization in the nucleus. This trend was reversed in high stiffness regions (~30 kPa), where decreased cellular and nuclear volumes and reduced mechanosensitive protein nuclear localization were observed. Interestingly, cells in soft regions exhibited enhanced osteogenic RUNX2 expression, while those in stiff regions upregulated the adipogenic regulator PPARγ, suggesting that volume, not substrate stiffness, is sufficient to drive 3D stem cell differentiation. Inhibition of myosin II (Blebbistatin) and ROCK (Y-27632), both key drivers of actomyosin contractility, resulted in reduced cell volume, especially in low stiffness regions, causing a decorrelation between volume expansion and mechanosensitive protein localization. Constitutively active and inactive forms of the canonical downstream mechanotransduction effector TAZ were stably transfected into ASCs. Activated TAZ resulted in higher cellular volume despite increasing stiffness and a consistent, stiffness-independent translocation of YAP and MRTFa into the nucleus. Thus, volume adaptation as a function of 3D matrix stiffness can control stem cell mechanotransduction and differentiation.",
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author = "Luke Major and Holle, {Andrew W} and Young, {Jennifer L} and Matt Hepburn and Kwanghee Jeong and Ian Chin and Rowan Sanderson and Jeong, {Ji Hoon} and Zach Aman and Brendan Kennedy and Yongsung Hwang and Dong-Wook Han and Park, {Hyun Woo} and Kun-Liang Guan and Spatz, {Joachim P} and Choi, {Yu Suk}",
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month = "12",
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Major, L, Holle, AW, Young, JL, Hepburn, M, Jeong, K, Chin, I, Sanderson, R, Jeong, JH, Aman, Z, Kennedy, B, Hwang, Y, Han, D-W, Park, HW, Guan, K-L, Spatz, JP & Choi, YS 2019, 'Volume adaptation controls stem cell mechanotransduction' ACS Applied Materials and Interfaces, vol. 11, no. 49, pp. 45520-45530. https://doi.org/10.1021/acsami.9b19770, https://doi.org/10.1021/acsami.9b19770

Volume adaptation controls stem cell mechanotransduction. / Major, Luke; Holle, Andrew W; Young, Jennifer L; Hepburn, Matt; Jeong, Kwanghee; Chin, Ian; Sanderson, Rowan; Jeong, Ji Hoon; Aman, Zach; Kennedy, Brendan; Hwang, Yongsung; Han, Dong-Wook ; Park, Hyun Woo; Guan, Kun-Liang; Spatz, Joachim P; Choi, Yu Suk.

In: ACS Applied Materials and Interfaces, Vol. 11, No. 49, 11.12.2019, p. 45520-45530.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Volume adaptation controls stem cell mechanotransduction

AU - Major, Luke

AU - Holle, Andrew W

AU - Young, Jennifer L

AU - Hepburn, Matt

AU - Jeong, Kwanghee

AU - Chin, Ian

AU - Sanderson, Rowan

AU - Jeong, Ji Hoon

AU - Aman, Zach

AU - Kennedy, Brendan

AU - Hwang, Yongsung

AU - Han, Dong-Wook

AU - Park, Hyun Woo

AU - Guan, Kun-Liang

AU - Spatz, Joachim P

AU - Choi, Yu Suk

PY - 2019/12/11

Y1 - 2019/12/11

N2 - Recent studies have found discordant mechanosensitive outcomes when comparing 2D and 3D, highlighting the need for tools to study mechanotransduction in 3D across a wide spectrum of stiffness. A gelatin methacryloyl (GelMA) hydrogel with a continuous stiffness gradient ranging from 5 to 38 kPa was developed to recapitulate physiological stiffness conditions. Adipose-derived stem cells (ASCs) were encapsulated in this hydrogel and their morphological characteristics and expression of both mechanosensitive proteins (Lamin A, YAP, and MRTFa) and differentiation markers (PPARγ and RUNX2) were analyzed. Low stiffness regions (~8 kPa) permitted increased cellular and nuclear volume and enhanced mechanosensitive protein localization in the nucleus. This trend was reversed in high stiffness regions (~30 kPa), where decreased cellular and nuclear volumes and reduced mechanosensitive protein nuclear localization were observed. Interestingly, cells in soft regions exhibited enhanced osteogenic RUNX2 expression, while those in stiff regions upregulated the adipogenic regulator PPARγ, suggesting that volume, not substrate stiffness, is sufficient to drive 3D stem cell differentiation. Inhibition of myosin II (Blebbistatin) and ROCK (Y-27632), both key drivers of actomyosin contractility, resulted in reduced cell volume, especially in low stiffness regions, causing a decorrelation between volume expansion and mechanosensitive protein localization. Constitutively active and inactive forms of the canonical downstream mechanotransduction effector TAZ were stably transfected into ASCs. Activated TAZ resulted in higher cellular volume despite increasing stiffness and a consistent, stiffness-independent translocation of YAP and MRTFa into the nucleus. Thus, volume adaptation as a function of 3D matrix stiffness can control stem cell mechanotransduction and differentiation.

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KW - cellular volume

KW - extracellular matrix

KW - mechanobiology

KW - mechanotransduction

KW - stem cell differentiation

KW - stiffness gradient

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U2 - 10.1021/acsami.9b19770

DO - 10.1021/acsami.9b19770

M3 - Article

VL - 11

SP - 45520

EP - 45530

JO - Applied Materials and Interfaces

JF - Applied Materials and Interfaces

SN - 1944-8244

IS - 49

ER -