Effect of geometrical constraints on human pluripotent stem cell nuclei in pluripotency and differentiation

Integrative Biology : Quantitative Biosciences From Nano to Macro
Eleonora GrespanNicola Elvassore

Abstract

Mechanical stimuli and geometrical constraints transmitted across the cytoskeleton to the nucleus affect the nuclear morphology and cell function. Human pluripotent stem cells (hPSCs) represent an effective tool for evaluating transitions in nuclear deformability from the pluripotent to differentiated stage, and for deciphering the underlying mechanisms. We report the first study that investigates the nuclear deformability induced by geometrical constraints of hPSCs both in the pluripotent stage and during early germ layer specification. We specifically developed micro-structured surfaces coupled with high-content imaging analysis algorithms to quantitatively characterize nuclear deformability. Our results show that hPSCs possess high nuclear deformability, which does not alter pluripotency. We observed nuclear deformability transition along early germ layer specification: during early ectoderm differentiation nuclear deformability is strongly reduced, during early endoderm differentiation nuclei keep a deformed shape and during early mesoderm specification they show an intermediate behaviour. Different mRNA expressions between hPSCs differentiated on flat and micro-structured surfaces have been observed along early mesoderm an...Continue Reading

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Citations

Mar 25, 2019·Stem Cells International·Rosaria SantoroGiulio Pompilio
Oct 28, 2019·Frontiers in Bioengineering and Biotechnology·Stephen LenziniJae-Won Shin
Aug 12, 2019·ACS Biomaterials Science & Engineering·Yan-Lei FanXi-Qiao Feng

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Methods Mentioned

BETA
confocal microscopy
Transfection
PCR

Software Mentioned

ABI Prism 7000 SDS
MATLAB

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