Figure - PMC

Skip to main content

An official website of the United States government

Here's how you know

Here's how you know

Official websites use .gov
A .gov website belongs to an official government organization in the United States.

Secure .gov websites use HTTPS
A lock ( ) or https:// means you've safely connected to the .gov website. Share sensitive information only on official, secure websites.

View full-text article in PMC

. 2019 Jun 17;9:8671. doi: 10.1038/s41598-019-45191-2

Search in PMC
Search in PubMed
View in NLM Catalog
Add to search

© The Author(s) 2019

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

PMC Copyright notice

Analysis of senescent ASC and dermal fibroblasts functional properties. (A) ASC adipogenic potential was assessed 7 days after ASC treatment (CAP or NT) by induction of differentiation (Diff.) and analysis by Oil-Red-O staining of lipid droplets (day 21 after induction of differentiation) and by RT-qPCR for adipogenic gene expression PPARγ2, LPL, Adiponectin and C/EBPα (day 14 after induction of differentiation) in comparison to ASC cultivated in control medium (Ctrl, no differentiation). Pictures were made with Nikon, Eclipse, TE2000-S microscope. RT-qPCR results are represented in fold increase to control medium condition (dot line) for the different genes (n = 4). (B) ASC were tested for their immunosuppression activity by co-cultivating ASC with activated T lymphocytes (TL) labelled with the CFSE marker. The percentage of immunosuppression is determined by flow cytometry after 5 days (proliferating TL = % TL CFSE negative) (n = 6). (C) Effect on macrophage polarization was performed by co-cultivating ASC with M0 macrophages (monocytes cultivated 6 days with M-CSF) and macrophage phenotype analysis by flow cytometry at indicated times for membrane markers CD45, HLA-DR and CD206. Data are represented in mean of fluorescent intensity (MFI) normalized to isotype control among living macrophages (DAPI negative and CD45 positive cells) (n = 4). (D) Dermal fibroblasts 7 days after treatment (CAP or NT) were tested for their differentiation into myofibroblast under TGF-β stimulation by RT-qPCR for αSMA, CTGF, vimentin, MMP1, fibronectin, collagen COL1a, COL3A1 and caldesmone (72 hours). Data are represented in fold increase to control media (n = 3). (E) Dermal fibroblast activation was also estimated by α-SMA labeling and imaging using the high content imaging system Operetta (Perkin Elmer, lens x20) (n = 4). Data represent mean ± SEM of independent experiments as indicated (n).