Mechanically and Chemically Tunable Cell Culture System for Studying the Myofibroblast Phenotype

Saums, Michele K; Wang, Weifeng; Han, Biao; Madhavan, Lakshmi; Han, Lin; Lee, Daeyeon; Wells, Rebecca G.

doi:10.1021/la4047758

Cited by 32 publications

(31 citation statements)

References 30 publications

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“…15, 25, 26 Therefore, for soft and stiff static hydrogels crosslinked with DTT only we targeted starting elastic moduli of ~ 15-20 kPa and ~ 1-3 kPa corresponding to diseased and healthy liver tissue, respectively. For the stiff-to-soft hydrogel we targeted initial mechanics of ~ 15-20 kPa with final mechanics of ~ 1-3 kPa after 14 days.…”

Section: Resultsmentioning

confidence: 99%

Gradually softening hydrogels for modeling hepatic stellate cell behavior during fibrosis regression

et al. 2016

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The extracellular matrix (ECM) presents an evolving set of mechanical cues to resident cells. We developed methacrylated hyaluronic acid (MeHA) hydrogels containing both stable and hydrolytically degradable crosslinks to provide cells with a gradually softening (but not fully degradable) milieu, mimicking physiological events such as fibrosis regression. To demonstrate the utility of this cell culture system, we studied the phenotype of rat hepatic stellate cells, the major liver precursors of fibrogenic myofibroblasts, within this softening environment. Stellate cells that were mechanically primed on tissue culture plastic attained a myofibroblast phenotype, which persisted when seeded onto stiff (~ 20 kPa) hydrogels. However, mechanically primed stellate cells on stiff-to-soft (~ 20 to ~ 3 kPa) hydrogels showed reversion of the myofibroblast phenotype over 14 days, with reductions in cell area, expression of the myofibroblast marker alpha-smooth muscle actin (α-SMA), and Yes-associated protein/Transcriptional coactivator with PDZ-binding motif (YAP/TAZ) nuclear localization when compared to stellate cells on stiff hydrogels. Cells on stiff-to-soft hydrogels did not fully revert, however. They displayed reduced expression of glial fibrillary acidic protein (GFAP), and underwent abnormally rapid re-activation to myofibroblasts in response to re-stiffening of the hydrogels through introduction of additional crosslinks. These features are typical of stellate cells with an intermediate phenotype, reported to occur in vivo with fibrosis regression and re-injury. Together, these data suggest that mechanics play an important role in fibrosis regression and that integrating dynamic mechanical cues into model systems helps capture cell behaviors observed in vivo.

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Section: Resultsmentioning

confidence: 99%

Gradually softening hydrogels for modeling hepatic stellate cell behavior during fibrosis regression

et al. 2016

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“…Culture on substrates produced with the stiffness of fibrotic scars activates a variety of different progenitors to become myofibroblasts (Hinz, 2010b;Saums et al, 2014). In fact, all fibroblastic cells are activated to form stress fibers by exposure to stiff (GPa) plastic dishes in standard culture and a characteristic fraction of these cells will spontaneously express α-SMA (Hinz, 2010b) (Figure 1).…”

Section: Matrix Mechanics and Cell Stressmentioning

confidence: 99%

Myofibroblasts

Hinz¹

2016

Experimental Eye Research

364

341

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Myofibroblasts are activated in response to tissue injury with the primary task to repair lost or damaged extracellular matrix. Enhanced collagen secretion and subsequent contraction - scarring - are part of the normal wound healing response and crucial to restore tissue integrity. Due to myofibroblasts ability to repair but not regenerate, accumulation of scar tissue is always associated with reduced organ performance. This is a fair price to pay by the body for not falling apart. Whereas myofibroblasts typically vanish after successful repair, dysregulation of the normal repair process can lead to persistent myofibroblast activation, for instance by chronic inflammation or mechanical stress in the tissue. Excessive repair leads to the accumulation of stiff collagenous ECM contractures - fibrosis - with dramatic consequences for organ function. The clinical need to terminate detrimental myofibroblast activities has stimulated researchers to answer a number of essential questions: where do myofibroblasts come from, what are the factors leading to their activation, how do we discriminate myofibroblasts from other cells, what is the molecular basis for their contractile activity, and how can we stop or at least control them? This article reviews the current state of the myofibroblast literature by emphasizing their role in ocular repair and fibrosis. It appears that although the eye is quite an extraordinary organ, ocular myofibroblasts behave or misbehave just like their siblings in other organs.

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“…Besides influencing cellular behavior based on specific molecular signaling, dimensionality and biomechanical cues in the lung clearly impact the effect of ECM on interstitial cell phenotype (8)(9)(10)(11)(12)(13)(14)(15). Although traditional in vitro studies of cell-matrix interactions usually rely on matrix-coated culture dishes, these artificial environments do not truly recapitulate the in vivo conditions under which cells reside.…”

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Lung Extracellular Matrix and Fibroblast Function

2015

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Extracellular matrix (ECM) is a tissue-specific macromolecular structure that provides physical support to tissues and is essential for normal organ function. In the lung, ECM plays an active role in shaping cell behavior both in health and disease by virtue of the contextual clues it imparts to cells. Qualities including dimensionality, molecular composition, and intrinsic stiffness all promote normal function of the lung ECM. Alterations in composition and/or modulation of stiffness of the focally injured or diseased lung ECM microenvironment plays a part in reparative processes performed by fibroblasts. Under conditions of remodeling or in disease states, inhomogeneous stiffening (or softening) of the pathologic ECM may both precede modifications in cell behavior and be a result of disease progression. The ability of ECM to stimulate further ECM production by fibroblasts and drive disease progression has potentially significant implications for mesenchymal stromal cell-based therapies; in the setting of pathologic ECM stiffness or composition, the therapeutic intent of progenitor cells may be subverted. Taken together, current data suggest that lung ECM actively contributes to health and disease; thus, mediators of cell-ECM signaling or factors that influence ECM stiffness may represent viable therapeutic targets in many lung disorders.

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Mechanically and Chemically Tunable Cell Culture System for Studying the Myofibroblast Phenotype

Cited by 32 publications

References 30 publications

Gradually softening hydrogels for modeling hepatic stellate cell behavior during fibrosis regression

Gradually softening hydrogels for modeling hepatic stellate cell behavior during fibrosis regression

Myofibroblasts

Lung Extracellular Matrix and Fibroblast Function

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