Human induced mesenchymal stem cells display increased sensitivity to matrix stiffness

Abstract

The clinical translation of mesenchymal stem cells (MSCs) is limited by population heterogeneity and inconsistent responses to engineered signals. Specifically, the extent in which MSCs respond to mechanical cues varies significantly across MSC lines. Although induced pluripotent stem cells (iPSCs) have recently emerged as a novel cell source for creating highly homogeneous MSC (iMSC) lines, cellular mechanosensing of iMSCs on engineered materials with defined mechanics is not well understood. Here, we tested the mechanosensing properties of three human iMSC lines derived from iPSCs generated using a fully automated platform. Stiffness-driven changes in morphology were comparable between MSCs and iMSCs cultured atop hydrogels of different stiffness. However, contrary to tissue derived MSCs, no significant changes in iMSC morphology were observed between iMSC lines atop different stiffness hydrogels, demonstrating a consistent response to mechanical signals. Further, stiffness-driven changes in mechanosensitive biomarkers were more pronounced in iMSCs than MSCs, which shows that iMSCs are more adaptive and responsive to mechanical cues than MSCs. This study reports that iMSCs are a promising stem cell source for basic and applied research due to their homogeneity and high sensitivity to engineered mechanical signals.

Manipulating Stem Cell Fate with Disordered Bioactive Cues on Surfaces: The Role of Bioactive Ligand Selection

“…Stem cells, known for their unique self-renewal and differentiation capabilities, are at the forefront of research interest. However, a significant challenge in the field remains the precise control over stem cell growth and differentiation. , The cellular behavior is profoundly influenced by the multifaceted extracellular matrix (ECM) environment, encompassing factors such as the ECM protein concentration and various stimuli from the stem cell niche. These stimuli include both chemical cues, ,,− such as composition ,,,,,− and ligand density, and physical factors like matrix stiffness, − surface topography, ,, and interfacial hydrophobicity .…”

A Polymer Canvas with the Stiffness of the Bone Matrix to Study and Control Mesenchymal Stem Cell Response

“…Cell Culture: Human iMSCs were derived and characterized as previously described. [5,43] For all studies, aliquots of cell suspension were seeded to the PMCC-coated ITO glasses placed in ultra-low attachment plates (Fisher Scientific) at a density of ≈1000-10 000 cells cm −2 . To study attachment and proliferation, cells were cultured in an expansion medium, consisting of high-glucose KnockOut Dulbecco's modified Eagle's medium (DMEM; Gibco) supplemented with 20% (v/v) HyClone fetal bovine serum (FBS; GE Healthcare Life Sciences), beta-fibroblast growth factor (1 ng mL −1 ; Invitrogen), nonessential amino acids (0.1 mm; Gibco), GlutaMAX (2 mm; Gibco), beta-mercaptoethanol (0.1 mm; Gibco), and antibiotic-antimycotic (100 U mL −1 ; Gibco), while to study osteogenic differentiation cells were cultured in the commercially available OsteoLife Complete Osteogenesis Medium (Lifeline Cell Technologies).…”

“…[1] In particular, tissue stiffness controls cell adhesion, migration, and proliferation, and acts as a strong driver for phenotypic commitment both in vivo and in vitro. [2][3][4][5] For example, atop soft hydrogels that mimic the stiffness of brain tissue, mesenchymal stem cells (MSCs) express neuronal biomarkers, whereas on rigid substrates they produce osteocalcin, a bone tissue-specific protein. [4] So far, most of the investigations focused on understanding the effect of stiffness on cell behavior and differentiation rely on substrates having stiffness values ranging from kPa to a few MPa.…”

Controlling differentiation of stem cells via bioactive disordered cues

“…Morphological results showed that the hMSC morphology generally tended to have an elongated and flattened shape, characterized by high anisotropy, high aspect ratio, and low circularity. 107,108 Most of the induced cells grew to be cuboidal in shape, showing low anisotropy, a low aspect ratio, and high circularity. 109,110 Few cells were induced to tend to the stellate morphology, exhibiting low anisotropy, a low aspect ratio, and moderate circularity.…”