(Top): Interdigitated electrode geometry used to determine the DEP cross-over frequency f xo. (Bottom): comsol Multiphysics simulation of the DEP force (log10[N]) acting on a 10 μm diameter cell (Clausius-Mossotti factor = 0.5) with an applied voltage of 3 V (pk), as a function of distance (x μm) along and height above (z μm) the electrode plane.
Immunocytochemical confirmation of hESC and derivative lineages. Undifferentiated H1 hESCs (a) and MSC-like cells (b), and RCM-1 trophoblast-like cells ((c), (d)) stained for OCT4 (a), MSC lineage markers CD105 (red) and Stro-1 (green) (b) and trophoblast markers CDX2 (green, (c)) and hCG-β (green, (d)). Nuclei are counterstained with DAPI (blue) in all panels. Scale bar equals to 20 μm.
Flow cytometric analysis of H1-MSC-like cells confirmed they were negative for CD14, CD34, and positive for CD105 (64%), CD90 (40%), CD73 (99%), and Stro1 (58%).
Differentiation potential of hyaluronan-generated hES-derived MSC-like cells. (a) H1HA cells differentiate along the ostoeogenic lineage to form mineralised bone when cultured in conditions conducive to osteogenesis (+OS) but not control conditions (−OS), as detected by alizarin red staining (red staining detectable from day 7 onwards). (b) Calcium deposition quantified by Sigma calcium assay kit from the experiment in (a) shows that mineralised calcium only accumulates to detectable levels in osteogenic conditions (+OS, black bars).
(a) Percentage distributions of the DEP cross-over frequencies (f xo) measured for the different human embryonic stem cell lines. (b) The derived membrane capacitance (C m) values.
Percentage distributions of the DEP cross-over frequencies (f xo) for (a) H1 and differentiated H1-MSC, (b) H9 and H9-MSC, (c) RCM1 and RCM1-trophoblast.
Membrane capacitance (C m) values obtained for the human embryonic stem cells and their differentiated progeny.
The progression of (a) mean membrane capacitance C m with 95% confidence interval bars, and (b) mean cell diameter with 95% confidence interval error and passage number for the H9 cells cultured on an hyaluronan coated substrate. (**** and *** signify a statistical difference with p < 0.0001 and p < 0.001 between groups).
Flow cytometry analysis data for five undifferentiated hESC lines used in the study confirmed that comparable and high level of detection of SSEA-4 and TRA-1-60 and low levels of SSEA-1, as expected for this phenotype. Passage number at which cells assessed denoted by “p.”
Summary of the data (cell diameter, DEP cross-over frequency (f xo), and membrane capacitance (C m)) obtained for the human embryonic stem cells and their differentiated progeny.
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