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      Stiffening hydrogels to probe short- and long-term cellular responses to dynamic mechanics.

      Nature Communications
      Biomechanical Phenomena, Cell Adhesion, Cell Culture Techniques, instrumentation, Cell Movement, Cells, Cultured, Extracellular Matrix, chemistry, Humans, Hydrogels, Mesenchymal Stromal Cells, cytology, Time Factors

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          Abstract

          Biological processes are dynamic in nature, and growing evidence suggests that matrix stiffening is particularly decisive during development, wound healing and disease; yet, nearly all in vitro models are static. Here we introduce a step-wise approach, addition then light-mediated crosslinking, to fabricate hydrogels that stiffen (for example, ~3-30 kPa) in the presence of cells, and investigated the short-term (minutes-to-hours) and long-term (days-to-weeks) cell response to dynamic stiffening. When substrates are stiffened, adhered human mesenchymal stem cells increase their area from ~500 to 3,000 μm(2) and exhibit greater traction from ~1 to 10 kPa over a timescale of hours. For longer cultures up to 14 days, human mesenchymal stem cells selectively differentiate based on the period of culture, before or after stiffening, such that adipogenic differentiation is favoured for later stiffening, whereas osteogenic differentiation is favoured for earlier stiffening.

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