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Dielectrophoresis based discrimination of human embryonic stem cells from differentiating derivatives
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10.1063/1.4771316
/content/aip/journal/bmf/6/4/10.1063/1.4771316
http://aip.metastore.ingenta.com/content/aip/journal/bmf/6/4/10.1063/1.4771316

Figures

Image of FIG. 1.
FIG. 1.

(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.

Image of FIG. 2.
FIG. 2.

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.

Image of FIG. 3.
FIG. 3.

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%).

Image of FIG. 4.
FIG. 4.

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).

Image of FIG. 5.
FIG. 5.

(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.

Image of FIG. 6.
FIG. 6.

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.

Image of FIG. 7.
FIG. 7.

Membrane capacitance (C m) values obtained for the human embryonic stem cells and their differentiated progeny.

Image of FIG. 8.
FIG. 8.

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).

Tables

Generic image for table
Table I.

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.”

Generic image for table
Table II.

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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/content/aip/journal/bmf/6/4/10.1063/1.4771316
2012-12-12
2014-04-18
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752b84549af89a08dbdd7fdb8b9568b5 journal.articlezxybnytfddd
Scitation: Dielectrophoresis based discrimination of human embryonic stem cells from differentiating derivatives
http://aip.metastore.ingenta.com/content/aip/journal/bmf/6/4/10.1063/1.4771316
10.1063/1.4771316
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