Biophysical subsets of embryonic stem cells display distinct phenotypic and morphological signatures

Abstract

The highly proliferative and pluripotent characteristics of embryonic stem cells engender great promise for tissue engineering and regenerative medicine, but the rapid identification and isolation of target cell phenotypes remains challenging. Therefore, the objectives of this study were to characterize cell mechanics as a function of differentiation and to employ differences in cell stiffness to select population subsets with distinct mechanical, morphological, and biological properties. Biomechanical analysis with atomic force microscopy revealed that embryonic stem cells stiffened within one day of differentiation induced by leukemia inhibitory factor removal, with a lagging but pronounced change from spherical to spindle-shaped cell morphology. A microfluidic device was then employed to sort a differentially labeled mixture of pluripotent and differentiating cells based on stiffness, resulting in pluripotent cell enrichment in the soft device outlet. Furthermore, sorting an unlabeled population of partially differentiated cells produced a subset of “soft” cells that was enriched for the pluripotent phenotype, as assessed by post-sort characterization of cell mechanics, morphology, and gene expression. The results of this study indicate that intrinsic cell mechanical properties might serve as a basis for efficient, high-throughput, and label-free isolation of pluripotent stem cells, which will facilitate a greater biological understanding of pluripotency and advance the potential of pluripotent stem cell differentiated progeny as cell sources for tissue engineering and regenerative medicine.

Cancer Stem Cell Plasticity – A Deadly Deal

“…Further, exogenous alteration of cell stiffness also resulted in changes in metastatic potential of these cells with less stiff cells showing greater invasion and migration (Chen et al, 2019). The results observed in this study corroborated well with earlier studies showing that cancer cells with greater deformability, lower adhesion strength and lower contractile force show enhanced metastatic potential (Swaminathan et al, 2011;Kraning-Rush et al, 2012;Byun et al, 2013;Sun et al, 2016;Bongiorno et al, 2018). Therefore, with increasingly detailed characterization of biomechanical properties of various subpopulations of cancer cells and ECM, a "mechanosome" or "matrisome" signature may be helpful in identifying and isolating the most aggressive cancer cell subpopulations (Roy Choudhury et al, 2019).…”

Cancer Stem Cell Plasticity – A Deadly Deal

“…Further, exogenous alteration of cell stiffness also resulted in changes in metastatic potential of these cells with less stiff cells showing greater invasion and migration (Chen, Allen et al 2019). The results observed in this study corroborated well with earlier studies showing that cancer cells with greater deformability, lower adhesion strength and lower contractile force show enhanced metastatic potential (Swaminathan, Mythreye et al 2011, Kraning-Rush, Califano et al 2012, Byun, Son et al 2013, Sun, Luo et al 2016, Bongiorno, Gura et al 2018). Therefore, with increasingly detailed characterization of biomechanical properties of various subpopulations of cancer cells and ECM, a 'mechanosome' or 'matrisome' signature may be helpful in identifying and isolating the most aggressive cancer cell subpopulations (Roy Choudhury, Gupta et al 2019).…”

Evaluating Oxygen Tensions Related to Bone Marrow and Matrix for MSC Differentiation in 2D and 3D Biomimetic Lamellar Scaffolds

“…However, the variation in the MSC morphology was observed on the 3D scaffold surface only in the cases of 5% and 12% O 2 tensions. At 5% O 2 , cell clumps existed, and this change might indicate the retention of stem cell phenotype since circular cell clumps are typically seen in undifferentiated embryonic cell culture [ 38 ]. Individual spindle-shaped cells at 12% O 2 might show the MSC differentiation.…”