Stem cells have the ability to differentiate into specific cell types. The two defining characteristics of a stem cell are perpetual self-renewal and the ability to differentiate into a specialized adult cell type. There are two major classes of stem cells: pluripotent that can become any cell in the adult body, and multipotent that are restricted to becoming a more limited population of cells. Cell sources, characteristics, differentiation and therapeutic applications are discussed. Stem cells have great potential in tissue regeneration and repair but much still needs to be learned about their biology, manipulation and safety before their full therapeutic potential can be achieved.
The niche in which stem cells reside and differentiate is a complex physico-chemical microenvironment that regulates cell function. The role played by three-dimensional physical contours was studied on cell progeny derived from mouse embryonic stem cells using microtopographies created on PDMS membranes. While markers of differentiation were not affected, the proliferation of heterogeneous mouse embryonic stem cell-derived progeny was attenuated by 15 μm-, but not 5 μm-high microprojections. This reduction was reversed by Rho kinase and myosin light chain kinase inhibition, which diminishes the tension generating ability of stress fibers. Purified cardiomyocytes derived from embryonic stem cells also showed significant blunting of proliferation and increased beating rates compared to cells grown on flat substrates. Thus, proliferation of stem cell-derived progeny appears to be regulated by microtopography through tension-generation of contractility in the third-dimension. These results emphasize the importance of topographic cues in the modulation of stem cell progeny behavior.
The goal was to test if the physical properties of the microenvironment control proliferation and activity of the mouse embryonic stem cell (mESC) progeny. Microtopographic features were fabricated by photolithography to create 15μm high projections spaced 80 or 500μm apart tetragonally in poly‐dimethyl‐silicone (PDMS) membranes. mESC differentiation began in hanging drops, followed by suspension culture, before the resulting embryoid bodies were disassociated and plated on either flat or the microtextured surfaces. The number of heterogeneous SC derivatives observed with phase microscopy was 60 ± 20% (n=3) on the 80μm microprojections compared to flat PDMS. Similar results were seen for pure cardiomyocytes derived using a puromycin resistant cassette incorporated into the NCX1 promoter. Only 43±12% (n=3) and 75±16% (n=5) of the cardiomyocytes were found on the 80μm and 500μm spaced microprojections, respectively, compared to the flat PDMS. The beating rate per minute of the cardiomyocytes was recorded by video microscopy and was 1.8±0.4 fold higher on the microprojections compared to the flat with surprising changes in coefficients of variance of 0.50 and 0.21 respectively (n=5). Results suggest that microtopography affects both expansion and beating characteristics of mESC progeny. T32HL007692 and HL 62426.
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