The response of human induced pluripotent stem cells to cyclic temperature changes explored by BIO-AFM

Abstract

AbstractHuman induced pluripotent stem cells (hiPSCs) are highly sensitive to extrinsic physical and biochemical signals from their extracellular microenvironments. In this study, we analyzed the effect of cyclic temperature changes on hiPSCs behaviors, especially by means of scanning force microscopy (BIO-AFM). The alternation in cellular mechanics, as well as the secretion and pattern of deposition of extracellular matrix (ECM) protein in hiPSCs were evaluated. The arrangement of the actin cytoskeleton changed with the variation of the temperature. The rearranged cytoskeleton architecture led to the subsequent changes in cell mechanics (Young's modulus of hiPSCs). With the exposure to the cyclic cold stimuli, an increase in the average surface roughness (Ra) and roughness mean square (RMS) was detected. This observation might be at least in part due to the upregulated secretion of Laminin α5 during repeated temporary cooling. The expression of pluripotent markers, NANOG and SOX2, was not impaired in hiPSCs, when exposed to the cyclic cold stimuli for 24 h. Our findings provide an insight into the effect of temperature on the hiPSC behaviors, which may contribute to a better understanding of the application of locally controlled therapeutic hypothermia. Graphic abstract The cyclic temperature changes, from 37 to 10 °C, rapidly increased the mechanical strength of human-induced pluripotent stem cells (hiPSCs), which could be explained by the re-arrangement of cytoskeletons. The capacity of hiPSCs to remodel the extracellular matrix was also altered by the repeated temporary cooling, as they exhibit an enhanced ability to physically remodulate and secrete the ECM components.