Substrate stiffness effect and chromosome missegregation in hIPS cells

Abstract

BackgroundEnsuring genetic stability in pluripotent stem cell (PSC) cultures is essential for the development of successful cell therapies. Although most instances lead to failed experiments and go unreported in the literature, many laboratories have found the emergence of genetic abnormalities in PSCs when cultured in vitro for prolonged amounts of time. These cells are primarily cultured in non-physiological stiff substrates like tissue culture polystyrene (TCPS) which raises the possibility that the cause of these abnormalities may be influenced by substrate mechanics.FindingsIn order to investigate this, human PSCs were grown on substrates of varying stiffness such as a range of polyacrylamide formulations, TCPS, and borosilicate glass coverslips. These substrates allowed for the testing of a stiffness range from 5kPa to 64GPa. Two human induced PSC (iPSC) lines were analyzed in this study: 19-9-11 iPSCs and 19.7 clone F iPSCs. Centrosome and DNA staining revealed that 19-9-11 iPSCs range from 1–8.5 % abnormal mitoses under the different culture conditions. A range of 4.4–8.1 % abnormal mitoses was found for 19.7 clone F iPSCs.ConclusionsAbnormal cell division was not biased towards one particular substrate. It was confirmed by Analysis of Variance (ANOVA) and Tukey’s Honest Significant Difference test that there was no statistically significant difference between passage numbers, cell lines, or substrates.Electronic supplementary materialThe online version of this article (doi:10.1186/s12952-015-0042-8) contains supplementary material, which is available to authorized users.

A bioengineered lymphatic vessel model for studying lymphatic endothelial cell‐cell junction and barrier function

“…After 1 day of culture, we fixed the cells and stained them with phalloidin and anti‐VE‐cad antibodies. On the plastic dish, in relatively stiffer condition with a Young's modulus of 2.28–3.28 GPa 64 compared to the Young's modulus of collagen 1 at only 0.5–12 kPa, 65 LECs appeared to have very tightened junctions. F‐actin and VE‐cad were strongly localized in the junctional area (Figure 2A, top).…”

Dystrophin Cardiomyopathies: Clinical Management, Molecular Pathogenesis and Evolution towards Precision Medicine

“…Several works examined the role of matrix rigidity on cell reprogramming [ 211 , 217 , 219 , 220 , 221 ], underlining how pluripotent stem cell differentiation and further maturation could be directed by substrate stiffness [ 206 , 207 , 211 , 217 , 222 ]. In particular, a positive effect on cardiac commitment has been demonstrated by growing and differentiating iPSCs on substrates with stiffness resembling that of healthy cardiac tissue [ 209 , 223 , 224 ].…”

Decoupling the role of chemistry and microstructure in hMSCs response to an osteoinductive calcium phosphate ceramic

“…This effect was observed on both flat and microstructured discs. Difference is stiffness between the two materials (2.28–3.28 GPa for tissue culture PS [ 64 ] and 1–3.45 for poly(D,L lactic acid) [ 65 ] depending on its molecular weight; ∼1.4 GPa for the molecular weight comparable to what was used here [ 66 ]) may have been a reason for this, as increasingly more studies show that substrate stiffness may affect cell attachment, proliferation and fate [ [67] , [68] , [69] ]. Obviously, differences in chemistry between the two polymers cannot be fully excluded either, although to the best of our knowledge, PLA has not been reported to have an intrinsic effect on the expression of the osteogenic genes tested here [ 70 , 71 ].…”