Product Applications View our CytoSoft eBrochure Here Mesenchymal Stem Cells (1) MSCs on soft substrates (1 kPa) had a higher expression of chondrogenic marker collagen-II and adipogenic marker lipoprotein lipase, less spreading, fewer stress fibers and lower proliferation rates than on hard substrates (15 kPa). Fibroblasts (2) PDGF-induced chemotaxis and migration of pulmonary fibroblasts precultured on stiff substrates (25 kPa) were significantly higher than those of cells precultured on soft gels (2 kPa). Endothelial Cells (3) Endothelial cell contraction-mediated neutrophil transmigration using an in vitro model of the vascular endothelium. Neutrophil transmigration increased with increasing substrate stiffness below the endothelium. Neural Stem Cells (4) NSCs proliferated on substrates below 10 kPa and exhibited maximal proliferation on 3.5 kPa surfaces. Neuronal differentiation was favored on the softest surfaces with EY < 1 kPa. Oligodendrocyte differentiation was favored on stiffer scaffolds >7 kPa. Astrocyte differentiation was only observed on <1 and 3.5 kPa surfaces and represented less than 2% of the total cell population. Tumor Cells (5) CCN1/CYR61 is highly regulated by stiffness in endothelial cells. Stiffness‐induced CCN1 activates β‐catenin nuclear translocation and signaling and that this contributes to upregulate N‐cadherin levels on the surface of the endothelium, increasing cancer cell metastasis. Cancer Stem Cells (6) Fibrotic rigidities found in pancreatic cancer tissues promote elements of EMT, including increases in vimentin expression, decreases in E-cadherin expression, nuclear localization of β-catenin, YAP and TAZ, changes in cell shape towards a mesenchymal phenotype and induction of chemoresistance. Mammary Epithelium (7) ECM stiffness alone induces malignant phenotypes but that the effect is completely abrogated when accompanied by an increase in basement-membrane ligands. Hepatic Stellate Cells (8) Higher matrix stiffness leads to transdifferentiation of hepatic stellate cells into fibrogenic, proliferative, and contractile myofibroblasts. Cardiomyocytes (9) Inverse correlation between the material stiffness and the amplitude of contraction of the cardiac tissue constructs by changing the modulus of the matrix using different compositions of PEG and fibrinogen. Smooth Muscle Cells (10) Elastic modulus between 448 and 5804 Pa regulated SMC cytoskeletal assembly in 3D, with cells in stiff matrices having a slightly higher degree of F-actin bundling after prolonged culture. Song Li, et al. The Effect of Matrix Stiffness on the Differentiation of Mesenchymal Stem Cells in Response to TGF-β. Biomaterials. 2011 Jun; 32(16): 3921–3930. Hasegawa Y, et al. Matrix stiffness regulates migration of human lung fibroblasts. Physiol Rep. 2017 May;5(9). Helim Aranda-Espinoza, et al. Endothelial cell substrate stiffness influences neutrophil transmigration via myosin light chain kinase-dependent cell contraction. Blood. 2011 Aug 11; 118(6): 1632–1640. Molly S. Shoichet, et al. The effect of substrate stiffness on adult neural stem cell behavior. Volume 30, Issue 36, December 2009, Pages 6867-6878. Sara Zanivan, et al. Tumor matrix stiffness promotes metastatic cancer cell interaction with the endothelium. EMBO J. 2017 Aug 15; 36(16): 2373–2389. Río Hernández, et al. Matrix stiffness induces epithelial–mesenchymal transition and promotes chemoresistance in pancreatic cancer cells. Oncogenesis volume 6, page e352 (2017). David J. Mooney, et al. Extracellular matrix stiffness and composition jointly regulate the induction of malignant phenotypes in mammary epithelium. Nature Materials volume 13, pages 970–978 (2014). Wells RG, et al. The role of matrix stiffness in hepatic stellate cell activation and liver fibrosis. J Clin Gastroenterol. 2005 Apr;39(4 Suppl 2):S158-61. DrorSeliktar, et al. Matrix stiffness affects spontaneous contraction of cardiomyocytes cultured within a PEGylated fibrinogen biomaterial. Acta Biomaterialia. Volume 3, Issue 1, January 2007, Pages 33-41. Andrew J.Putnam, et al. The effects of matrix stiffness and RhoA on the phenotypic plasticity of smooth muscle cells in a 3-D biosynthetic hydrogel system. Biomaterials, volume 29, Issue 17, June 2008, Pages 2597-2607.

CytoSoft™ products provide a tool to culture cells on PDMS substrates with various rigidity covering a broad physiological range, including 0.2, 0.5, 2, 8, 16, 32, and 64 kPa. This CytoSoft™ product comes with 10 standard T-25 flasks with substrates of a chosen stiffness (selected below).

Product Description This CytoSoft™ T-25 Flask product has a defined elastic modulus (see certificate of analysis) in the bottom of a standard T-25 Flask. The thickness of the silicone gel is uniform with a ~0.5 mm thick layer of silicone in each flask that is fully compatible with mammalian cell cultures. The silicone gels are activated and ready to bind to a purified ECM, such as PureCol ® type I collagen (#5005) prior to cell addition. The plates are packaged sterilized, and provided with 10 flasks per package. The rigidity of the substrate to which cells adhere can have a profound effect on cell morphology and gene expression. CytoSoft™ products provide a tool to culture cells on substrates with various rigidities covering a broad physiological range. On the bottom of each flask, there is a thin layer of specially formulated biocompatible silicone, whose elastic modulus (rigidity) is carefully measured and certified. The surfaces of the gels in CytoSoft™ products are functionalized to form covalent bonds with amines on proteins. This chemical functionalization is stable and the reaction does not require a catalyst, facilitating the coating of the gel surfaces with matrix proteins and plating cells. The silicone substrates of CytoSoft™ products are optically clear and have a low auto-florescence. The layer of silicone in each flask is firmly bonded to the bottom of the flask. Unlike hydrogels (such as polyacrylamide gels), silicone gels are not susceptible to hydrolysis, do not dry nor swell, are resilient and resistant to tearing or cracking, and their elastic moduli (rigidities) remain nearly unchanged during extended storage periods. CytoSoft™ products accommodate the harvesting of cells using enzymes such as trypsin and collagenase. There is no biochemical breakdown of the substrate during or after enzyme treatment, and there are no residuals of the substrate in the sample retrieved from a CytoSoft™ plate. Parameter, Testing, and Method CytoSoft™ T-25 Flasks Sterilization Method Ozone Plate Size T-25 Quantity per Package 10 Flasks Rigidty (Elastic Modulus) 0.2, 0.5, 2, 8, 16, 32, 64 kPa (exact values provided on the CofA) Storage Temperature Room Temperature Shelf Life Minimum of 6 months from date of receipt Plate Surface Material Functionalized Silicone Growth Area per Well 25 cm 2 Cell Attachment Assay Pass