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1.
Figure 6

Figure 6. From: Clonal Populations of Amniotic Cells by Dilution and Direct Plating: Evidence for Hidden Diversity.

Immunostaining of fibronectin and N-cadherin. BMMSCp5, PB4A2p16, PB3B5p11, and PB1C4p12 cell populations were immunostained for fibronectin (grayscale), N-cadherin (red), and fluorescent chromatin dye (blue). Scale bar, 50 microns, is identical in all panels.

Patricia G. Wilson, et al. Stem Cells Int. 2012;2012:485950.
2.
Figure 7

Figure 7. From: Clonal Populations of Amniotic Cells by Dilution and Direct Plating: Evidence for Hidden Diversity.

Immunostaining of keratin, vimentin, and fibronectin in mixed cell populations. Mixed cell populations (a, e) ChM1, (b) ChM2, (c, f) ChM3, and (d) ChM4 were immunostained with antibodies against (a–f) keratins (red), (a–d) vimentin or (e, f) fibronectin (green) and (a–f) a fluorescent chromatin dye (blue). Insets in (a–d) show images of keratin signal at low magnification (0.3x). Scale bar, (a–d) 50 microns, (e, f) 100 microns.

Patricia G. Wilson, et al. Stem Cells Int. 2012;2012:485950.
3.
Figure 4

Figure 4. From: Clonal Populations of Amniotic Cells by Dilution and Direct Plating: Evidence for Hidden Diversity.

Adipogenic differentiation. Populations of BMMSCp5, PB4A2p19, PB1C4p11, and PB3B5p14 cells were maintained in adipogenic media for 3 to 4 weeks. Representative bright field images show robust adipogenic differentiation in BMMSCs and PB1C4 cell populations as indicated by oil red-O stained spheres. Arrows and inserts indicate very small oil red-O positive spheres in BMMSC, PB4A2, and PB1C4 populations that were not detected in PB3B5 populations. Magnification is identical in all panels.

Patricia G. Wilson, et al. Stem Cells Int. 2012;2012:485950.
4.
Figure 5

Figure 5. From: Clonal Populations of Amniotic Cells by Dilution and Direct Plating: Evidence for Hidden Diversity.

Immunostaining for vimentin and keratin. BMMSCp5, PB4A2p16, PB1C4p12 and PB3B5p11 cell populations were stained for the stromal cell marker vimentin (green), the epithelial cell marker keratin (red) and a fluorescent chromatin dye (blue). Note that virtually all PB4A2 and PB3B5 cells showed keratin staining, although the intensity varied. The clonal population of PB1C4 stromal progenitors did not show immunostaining of prominent networks of either vimentin or keratin. Scale (50 microns) is identical in all panels.

Patricia G. Wilson, et al. Stem Cells Int. 2012;2012:485950.
5.
Figure 3

Figure 3. From: Clonal Populations of Amniotic Cells by Dilution and Direct Plating: Evidence for Hidden Diversity.

Osteogenic differentiation with companion control populations. Populations of BMMSCp5, PB4A2p19, PB1C4p11, and PB3B5p14 cells were expanded in growth media and then maintained in osteogenic media for 3 to 4 weeks. Robust deposition of calcium, which stains with alizarin red, was generated in BMMSCs and PB4A2 cell populations, but not in PB1C4 or PB3B5 cell populations. The corresponding populations in control media without differentiation supplements are shown in the column on the right. Note the cross-hatched appearance of overly confluent populations of BMMSCp5, PB4A2p19, and PB1C4p11 cells while PB3B5p14 cell populations that apparently ceased proliferation near confluence. Magnification is identical in all panels.

Patricia G. Wilson, et al. Stem Cells Int. 2012;2012:485950.
6.
Figure 2

Figure 2. From: Clonal Populations of Amniotic Cells by Dilution and Direct Plating: Evidence for Hidden Diversity.

Flow cytometry of cell surface markers. Cell populations BMMSCp5, PB4A2p20, PB3B5p14, and PB1C4p15 were stained with antibodies against (a) CD73, CD29, CD44, SSEA4 and (b) against CD90 and CD105. Cell populations are indicated in vertical text on the left of the corresponding row of histograms for each marker. The markers and conjugated fluarochromes are indicated at the bottom of the corresponding column of each histogram. The x-axis of all histograms corresponds to the mean florescence intensity (MFI) in log scale. The y-axis is the percentage (%) of events in linear scale that were detected at each position of MFI along the x-axis. Cell populations were gated to exclude presumptive dead cells and debris. The histogram of isotype controls is depicted by black line and the marker that is assayed is depicted in the filled histogram. 10,000 events were scored for all populations. The percentage of immunostained cells, excluding those stained by isotype controls, is indicated within each histogram.

Patricia G. Wilson, et al. Stem Cells Int. 2012;2012:485950.
7.
Figure 1

Figure 1. From: Clonal Populations of Amniotic Cells by Dilution and Direct Plating: Evidence for Hidden Diversity.

Clonal population of cells from amniocentesis samples. Uncultured amniocentesis samples were diluted into growth media and plated into 24-well tissue-culture-treated plates. (a) Low-magnification image shows an expanding clonal population in one well of 24-well plate as a spherical cluster that is located near the well edge. Arrow indicates cells. (b) Higher magnification (5x) of the same population shows apparently well isolated cells. Representative clonal populations are shown here with passage numbers (p) indicated: (c) PB4A2p4, (d) PB3A5p3, (e) PB1C4p11, and (f) BMMSCp5 control cells. Stromal (c) PB4A2p4 and (e) PB1C4p11 cell populations resemble (f) BMMSCp4 cells, showing irregular cell boundaries while (d) PB3B5 epithelial cells were typically spheroid and often found in clusters with closely apposed boundaries. Scale bar, 100 microns. Magnification is identical in (c), (d), (e), and (f).

Patricia G. Wilson, et al. Stem Cells Int. 2012;2012:485950.

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