Stem cell migration and mechanotransduction on linear stiffness gradient hydrogels

Hadden, William J.; Young, Jennifer L.; Holle, Andrew W.; McFetridge, Meg L.; Kim, Du Yong; Wijesinghe, Philip; Taylor‐Weiner, Hermes; Wen, Jessica H.; Lee, Andrew; Bieback, Karen; Vo, Ba-Ngu; Sampson, David D.; Kennedy, Brendan F.; Spatz, Joachim P.; Engler, Adam J.; Choi, Yu Suk

doi:10.1073/pnas.1618239114

Cited by 410 publications

(353 citation statements)

References 68 publications

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“…Expression changes in mechanosensitive proteins verified typical ASC mechanotransduction occurring in response to ECM stiffness. Increased nuclear localization of YAP was observed on stiffer hydrogels (Figure b) in agreement with previous findings (Dupont et al, ; Hadden et al, ; Swift et al, ), and MRTF‐A followed the same trend (Figure b; p < 0.05) as expected from previous findings (Shiwen et al, ).…”

Section: Resultssupporting

confidence: 92%

Developing a high-throughput platform to direct adipogenic and osteogenic differentiation in adipose-derived stem cells

Major

Choi

2018

J Tissue Eng Regen Med

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The popular reductionist approach towards stem cell differentiation has identified an array of factors that can induce lineage-specific differentiation. Whether these variables direct differentiation in a multivariable setting in the same way as a single-variable setting is a question of interest. Polyacrylamide hydrogels of specific stiffness were microcontact printed with fibronectin in specific shapes and sizes to construct 54 different extracellular matrix types of defined stiffness, shape, and area. Three media types were also used to investigate the effect of both biomechanical and biochemical inducers of differentiation on adipose-derived stem cells. Stiffness was found to be a significant regulator of both adipogenic (3 kPa) and osteogenic (35 kPa) differentiation across all conditions. Biochemically induced osteogenic differentiation occurred as well as increased osteogenesis on larger extracellular matrix areas (above 5,000 μm ). The absence of clear trends for all variables demonstrates the atypical expression patterns that arise when variables that may work competitively or synergistically are considered together in a single, high-throughput system.

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Section: Resultssupporting

confidence: 92%

Developing a high-throughput platform to direct adipogenic and osteogenic differentiation in adipose-derived stem cells

Major

Choi

2018

J Tissue Eng Regen Med

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“…Durotaxis is a class of externally directed cell migration in which the direction of cell migration is governed by a local physical parameter: the stiffness of the migratory substrate (11)(12)(13)(14)(15)(16)(17)(18)(19)(20). Durotaxis has been characterized in detail recently in several nonmalignant cell types, generally those derived from a mesenchymal lineage (e.g., see (48,49)). However, it was not clear whether cancer cells can also undergo effective durotaxis.…”

Section: Discussionmentioning

confidence: 99%

Durotaxis by Human Cancer Cells

DuChez

Doyle

Dimitriadis

et al. 2019

Biophysical Journal

168

153

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Durotaxis is a type of directed cell migration in which cells respond to a gradient of extracellular stiffness. Using automated tracking of positional data for large sample sizes of single migrating cells, we investigated 1) whether cancer cells can undergo durotaxis; 2) whether cell durotactic efficiency varies depending on the regional compliance of stiffness gradients; 3) whether a specific cell migration parameter such as speed or time of migration correlates with durotaxis; and 4) whether Arp2/3, previously implicated in leading edge dynamics and migration, contributes to cancer cell durotaxis. Although durotaxis has been characterized primarily in nonmalignant mesenchymal cells, little is known about its role in cancer cell migration. Diffusible factors are known to affect cancer cell migration and metastasis. However, because many tumor microenvironments gradually stiffen, we hypothesized that durotaxis might also govern migration of cancer cells. We evaluated the durotactic potential of multiple cancer cell lines by employing substrate stiffness gradients mirroring the physiological stiffness encountered by cells in a variety of tissues. Automated cell tracking permitted rapid acquisition of positional data and robust statistical analyses for migrating cells. These durotaxis assays demonstrated that all cancer cell lines tested (two glioblastoma, metastatic breast cancer, and fibrosarcoma) migrated directionally in response to changes in extracellular stiffness. Unexpectedly, all cancer cell lines tested, as well as noninvasive human fibroblasts, displayed the strongest durotactic migratory response when migrating on the softest regions of stiffness gradients (2-7 kPa), with decreased responsiveness on stiff regions of gradients. Focusing on glioblastoma cells, durotactic forward migration index and displacement rates were relatively stable over time. Correlation analyses showed the expected correlation with displacement along the gradient but much less with persistence and none with cell speed. Finally, we found that inhibition of Arp2/3, an actin-nucleating protein necessary for lamellipodial protrusion, impaired durotactic migration.For all live-cell fluorescence experiments, DMEM without phenol red or FluoroBrite DMEM (GIBCO) were used, with each supplemented with a 1:100 ratio of Oxyfluor (Oxyrase, Mansfield, OH) and 10 mM DL-lactate (Sigma, St. Louis, MO) to reduce photobleaching and phototoxicity, Durotaxis by Human Cancer Cells

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“…Various techniques, including photochemical reactions with spatially controlled UV irradiation (Nemir, Hayenga, & West, ; Norris, Tseng, & Kasko, ; O'Connell et al, ; Sunyer, Jin, Nossal, & Sackett, ; Wong, Velasco, Rajagopalan, & Pham, ), physical crosslinking (Kim et al, ; Oh, An, Kim, & Lee, ), and diffusion‐based crosslinking (Hadden et al, ; Hartman, Isenberg, Chua, & Wong, ; Lee et al, ), have been suggested to realize the stiffness gradient platforms. The approaches were generally based on synthetic polymers or chemically modified natural polymers as substrate material, including polydimethylsiloxane (Wang, Tsai, & Voelcker, ), polyacrylamide (Hadden et al, ; Hopp et al, ; Sunyer et al, ), polyvinyl alcohol (Kim, An, et al, ; Oh et al, ), and gelatin (O'Connell et al, ). The stiffness gradient platforms enabled high‐throughput examinations of stiffness effect on various cellular responses in a certain range of the stiffness on a single substrate (Hopp et al, ; Kim, An, et al, ; Oh et al, ; Wang, Tsai, & Voelcker, ).…”

Section: Introductionmentioning

confidence: 99%

Grayscale mask‐assisted photochemical crosslinking for a dense collagen construct with stiffness gradient

et al. 2019

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Despite the potential of a collagen construct with a stiffness gradient for investigating cell–extracellular matrix (ECM) stiffness interaction or recapitulating an in vivo tissue interface, it has been developed in a limited way due to the low and poorly controllable mechanical properties of the collagen. This study proposes a novel fabrication process to achieve a compressed collagen construct with a stiffness gradient, named COSDIENT, at a level of ~ 1 MPa while maintaining in vivo ECM‐like dense collagen fibrillar structures. The COSDIENT was fabricated by collagen compression followed by grayscale mask‐assisted UV–riboflavin crosslinking. The collagen compression process enabled the remarkable increase in the stiffness of the collagen gel from ~ 1–10 kPa to ~ 1 MPa by physical compaction. The subsequent UV–riboflavin crosslinking with a continuous‐tone grayscale mask could simply generate a gradual change of UV irradiation followed by modulating riboflavin‐mediated crosslinking, thereby resulting in a continuous stiffness gradient with a range of 1.16–4.38 MPa in the single compressed collagen construct. The suggested grayscale mask‐assisted photochemical crosslinking had no effect on the physical and optical properties of the original compressed collagen construct, while inducing gradual changes of chemical bonds among collagen fibrils. A skin wound healing assay with epidermal keratinocytes was finally applied as an application example of the COSDIENT to examine the effect of stiffness on the skin keratinocyte behavior.

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Stem cell migration and mechanotransduction on linear stiffness gradient hydrogels

Cited by 410 publications

References 68 publications

Developing a high-throughput platform to direct adipogenic and osteogenic differentiation in adipose-derived stem cells

Developing a high-throughput platform to direct adipogenic and osteogenic differentiation in adipose-derived stem cells

Durotaxis by Human Cancer Cells

Grayscale mask‐assisted photochemical crosslinking for a dense collagen construct with stiffness gradient

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