Further characterization of embryonic stem cell-derived radial glial cells
Previously, we showed that radial glia‐like (RG) cells differentiated from embryonic stem (ES) cells after retinoic acid induction (Liour and Yu, 2003: Glia 42:109–117). In the present study, we demonstrate that the production of RG cells from ES cells is independent of the neural differentiation pr...
Saved in:
Published in | Glia Vol. 53; no. 1; pp. 43 - 56 |
---|---|
Main Authors | , , , , , |
Format | Journal Article |
Language | English |
Published |
Hoboken
Wiley Subscription Services, Inc., A Wiley Company
01.01.2006
Wiley-Liss |
Subjects | |
Online Access | Get full text |
Cover
Loading…
Summary: | Previously, we showed that radial glia‐like (RG) cells differentiated from embryonic stem (ES) cells after retinoic acid induction (Liour and Yu, 2003: Glia 42:109–117). In the present study, we demonstrate that the production of RG cells from ES cells is independent of the neural differentiation protocol used. These ES cell‐derived RG (ES‐RG) cells are similar in morphology to RG cells in vivo and express several characteristic RG cell markers. The processes of these ES‐RG cells are organized into radial arrays similar to the RG scaffold in developing CNS. Expression of Pax6, along with other circumstantial data, suggests that at least some of these ES‐RG cells are neural progenitors. The progression of neurogenesis into gliogenesis during the in vitro neural differentiation of ES cells recapitulates the in vivo developmental process. The identification of two cell surface markers, SSEA‐1 and GM1, on both the native embryonic RG cells and ES‐RG cells, may facilitate purification of radial glial cells for future studies and cell therapy. Overall, our study suggests that differentiation of radial glial cells is a common pathway during the neural differentiation of ES cells. © 2005 Wiley‐Liss, Inc. |
---|---|
Bibliography: | Morphology of three-dimensional rendering of a radial array at EFd11 which had been stained using an anti-nestin antibody (shown in green). A composite rendering of a Z-scan series of confocal images is shown in which the step size between each Z slice is 2.68 μm. The image is rendered from total 8 slices of image using Volovity v2.6.1 program (Improvision Ltd.). The image can be viewed using QuickTime Player software (available at http://www.apple.com/quicktime/download ), and rotated to view array from multiple angles revealing that each array is organized into approximately 3 cell layers containing long radially aligned nestin-positive processes. The scale of X-Y dimension is 500x500 μm.Higher magnification photos of ES cell-derived radial glial cells showing double-labeling of D3 ES cell cultures differentiated using the lineage selection protocol at EFd12 (A-C) or E14 cortex (D) using antibodies against vimentin, GLAST, SSEA-1 and nestin, or FITC-conjugated cholera toxin B subunit, as indicated by the color of the marker labels. Scale bars: 5 μm in A, B, and C or 20 μm in D.Merged image of Figure 1B showing that nestin and doublecortin immunolabeling is mutually exclusive in 2B6 cultures after differentiation using the lineage selection protocol. Similar results were seen with other ES cell lines and other differentiation protocols.Supporting Information file jws-glia.20257.lgd.doc istex:96CC074D07D8ACD99BD5E3684A5E7256E9203252 ArticleID:GLIA20257 Children's Medical Research Foundation National Institutes of Health - No. WS1185 ark:/67375/WNG-PVB9FKK8-3 ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 23 |
ISSN: | 0894-1491 1098-1136 |
DOI: | 10.1002/glia.20257 |