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Tissue-Specific Stem Cells
Article first published online: 5 JUL 2013
Copyright © 2013 AlphaMed Press
Volume 31, Issue 7, pages 1309–1320, July 2013
How to Cite
Gil-Ortega, M., Garidou, L., Barreau, C., Maumus, M., Breasson, L., Tavernier, G., García-Prieto, C. F., Bouloumié, A., Casteilla, L. and Sengenès, C. (2013), Native Adipose Stromal Cells Egress from Adipose Tissue In Vivo: Evidence During Lymph Node Activation. STEM CELLS, 31: 1309–1320. doi: 10.1002/stem.1375
Author contributions: M G-O., L.G., and C.S.: conception and design, collection and/or assembly of data, data analysis and interpretation, manuscript writing, and final approval of manuscript; C.B., M.M., L.B., G.T., and C.F.G-P.: collection and/or assembly of data and final approval of manuscript; A.B.: financial support, data analysis and interpretation, and final approval of manuscript; L.C.: conception and design, financial support, data analysis and interpretation, manuscript writing, and final approval of manuscript.
Disclosure of potential conflicts of interest is found at the end of this article.
first published online in STEM CELLS EXPRESS March 26, 2013.
- Issue published online: 5 JUL 2013
- Article first published online: 5 JUL 2013
- Accepted manuscript online: 26 MAR 2013 08:23AM EST
- Manuscript Accepted: 13 FEB 2013
- Manuscript Received: 9 NOV 2012
- Inserm, “La ligue nationale contre le cancer”
- Association Française d'étude et de recherches sur l'obésité
Additional Supporting Information may be found in the online version of this article.
|sc-12-1058_sm_SupplFigure1.pdf||228K||Figure S1. Sca-1+/CD45−/CD31- and CD45+ cell proportions in different fat depots. (A-C) Percentage or cell number per fat depot (D) of Sca-1+/CD45−/CD31- and CD45+ cells in the SVF from ScAT (A), PGAT (B) and MesAT (C). Results are expressed as mean ± SEM of n≥10 separate experiments. * p< 0.01 and ** p<0.001 when compared to ScAT. (E) Representative image of lipid droplet staining with BODIPY Dye in CD45+ cells after 10 days in adipogenic culture conditions.|
|sc-12-1058_sm_SupplFigure2.tif||1491K||Figure S2. AMD3100 treatment does not modify murine ASC viability and phenotype. (A) Percentage of dead cells in the SVF from SCAT, PGAT and MesAT treated or not with AMD3100 (10−6M) in vitro for 180 min. (B) Percentage of Sca-1+/CD45−/CD31- cells in the SVF from SCAT, PGAT and MesAT treated or not with AMD3100 (10−6M) in vitro for 180 min.|
|sc-12-1058_sm_SupplFigure3.tif||1664K||Figure S3. In vivo, the administration of CXCR4 antagonist AMD3100 does not increase the number of Sca-1+/CD45−/CD31- cells in blood. (A-C) Representative dot blots showing flow cytometry cellular staining profiles of blood samples from 10 week-old C57BL/6 mice. (A) Forward-scattered light (FSC) is proportional to cell-surface area or size and side-scattered light (SSC) is proportional to cell granularity or internal complexity. (B, C) Representative dot blots showing flow cytometry staining profiles into the CD45- cell population with Sca-1 and CD31 antibodies in mice treated (C) or not (B) avec AMD3100.|
|sc-12-1058_sm_SupplFigure4.pdf||291K||Figure S4. Lymph node structure and injected GFP+-ASCs localization. (A) Representative image of PLNs from mice injected in the footpad with GFP+ ASCs (in green) 12 hours before sacrifice and stained Hoescht 33342 (in blue). (B-D) Representative images of PLNs slices stained with (B) anti-B220 (red), (C) anti-CD3 (red) or anti-ER-TR7 (red) and Hoescht 33342 (blue). Representative microscopy results of at least 3 independent experiments.|
|sc-12-1058_sm_SupplFigure5.pdf||335K||Figure S5. T cell activation markers increase in OVA-IFA immunized draining lymph node while no changes are detected in BM cell content. Mice were immunized subcutaneously with 50μg of ovalbumin (OVA) in IFA in the right footpad. Left footpad was injected with saline. Nine days later, draining lymph node, popliteal (right) and inguinal (left) were separately harvested. (A) Lymph node cells were labelled with antibody cocktail. CD44 and CD62L expression gated on TCRb+CD4+ cells and CD44 and CD62L expression gated on TCRb+CD8+ cells. (B,C) The femurs from control or vaccinated hindlimbs were harvested and BMs were recovered by flushing. Total cell number was determined (B) (see methods) and the Sca- 1+/CD45−/CD31- cell content was determined by flow cytometry and compared to the one of control femurs, 9 days after the immunization procedure. Results are expressed as mean ± SEM of n=6 separate experiments.|
|sc-12-1058_sm_SupplVideo1.avi||52228K||Supplementary Video 1|
|sc-12-1058_sm_SupplVideo2.avi||2355K||Supplementary Video 2|
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