Intermittent compressive force induces cell cycling and reduces apoptosis in embryoid bodies of mouse induced pluripotent stem cells

Abstract

In vitro manipulation of induced pluripotent stem cells (iPSCs) by environmental factors is of great interest for three-dimensional (3D) tissue/organ induction. The effects of mechanical force depend on many factors, including force and cell type. However, information on such effects in iPSCs is lacking. The aim of this study was to identify a molecular mechanism in iPSCs responding to intermittent compressive force (ICF) by analyzing the global gene expression profile. Embryoid bodies of mouse iPSCs, attached on a tissue culture plate in 3D form, were subjected to ICF in serum-free culture medium for 24 h. Gene ontology analyses for RNA sequencing data demonstrated that genes differentially regulated by ICF were mainly associated with metabolic processes, membrane and protein binding. Topology-based analysis demonstrated that ICF induced genes in cell cycle categories and downregulated genes associated with metabolic processes. The Kyoto Encyclopedia of Genes and Genomes database revealed differentially regulated genes related to the p53 signaling pathway and cell cycle. qPCR analysis demonstrated significant upregulation of Ccnd1, Cdk6 and Ccng1. Flow cytometry showed that ICF induced cell cycle and proliferation, while reducing the number of apoptotic cells. ICF also upregulated transforming growth factor β1 (Tgfb1) at both mRNA and protein levels, and pretreatment with a TGF-β inhibitor (SB431542) prior to ICF abolished ICF-induced Ccnd1 and Cdk6 expression. Taken together, these findings show that TGF-β signaling in iPSCs enhances proliferation and decreases apoptosis in response to ICF, that could give rise to an efficient protocol to manipulate iPSCs for organoid fabrication.

Application of Shear Stress for Enhanced Osteogenic Differentiation of Mouse Induced Pluripotent Stem Cells

“…Mechanical stimulation has been reported to affect the growth and differentiation of stem cells. Our group previously demonstrated that intermittent compressive force promotes proliferation and suppresses apoptosis in RA-treated embryoid bodies of mouse iPSCs 21 . This implies a role for mechanical stimuli in promoting iPSC proliferation.…”

Application of shear stress for enhanced osteogenic differentiation of mouse induced pluripotent stem cells

“…Mechanical stimulation has been reported to affect the growth and differentiation of stem cells. Our group previously demonstrated that intermittent compressive force promotes proliferation and suppresses apoptosis in RA-treated embryoid bodies of mouse iPSCs 18 . This implies a role for mechanical stimuli in promoting iPSC proliferation.…”

Periostin‐integrin interaction regulates force‐induced TGF‐β1 and α‐SMA expression by hPDLSCs

“…An intermittent compressive force (ICF) was applied for 24 h. A compressive force apparatus was used, as described previously (Manokawinchoke et al, 2015; Manokawinchoke, Limraksasin, et al, 2022). The compressive force was generated by applying load on the culture medium using a pestle moving up and down in a matched‐size well plate.…”