Journal: Open Life Sciences

Article Title: TGF-β1-overexpressing mesenchymal stem cells reciprocally regulate Th17/Treg cells by regulating the expression of IFN-γ

doi: 10.1515/biol-2021-0118

Figure Lengend Snippet: The effect of TGF-β1/MSCs on T cells. (a) T cells were co-cultured with MSCs or TGF-β1/MSCs, and the proportion of CD4 + CD25 + T cells in lymphocytes was detected by flow cytometry. (b) CD4 + T cells were co-cultured with MSCs or TGF-β1/MSCs for 3, 5, or 7 days, and proliferative responses were measured. (c) CD8 + T cells were co-cultured with MSCs or TGF-β1/MSCs in the presence of IL-2 after 3, 5, or 7 days. Live cells were collected, and the cytotoxic activity against MDCK cells was assessed. Data are expressed as the mean ± SD. Three independent experiments were analyzed using unpaired Student’s t -tests. * P < 0.05; ** P < 0.01; MSCs group vs control group. # P < 0.05; ## P < 0.01; TGF-β1/MSCs group vs MSCs group.

Article Snippet: Fluorescence-labeled monoclonal antibodies anti-CD4, anti-CD25, anti-CD34, anti-CD105, anti-CD44, anti-CD29, and anti-CD90 were, respectively, added and incubated in the refrigerator at 4°C for 30 min. All antibodies for flow cytometry were purchased from BD Biosciences.

Techniques: Cell Culture, Flow Cytometry, Activity Assay

Journal: Open Life Sciences

Article Title: TGF-β1-overexpressing mesenchymal stem cells reciprocally regulate Th17/Treg cells by regulating the expression of IFN-γ

doi: 10.1515/biol-2021-0118

Figure Lengend Snippet: The effect of TGF-β1/MSCs on Th17 differentiation of T cells. (a) After CD4 + CD25 − T cells were cultured with TGF-β1/MSCs for 3 days in the presence of IL-2, cytokines (IL-4, IL-10, IL-17A, IL-21, and IL-22) were measured by ELISA from the culture supernatant. (b) The expression of IL17, Foxp3, and RORγt in T cells was measured by Western blotting. (c) The mRNA levels of T-bet, IL-17, IL-17F, RORγt, Foxp3, and RORα were detected using RT-PCR. Data are expressed as the mean ± SD. Three independent experiments were analyzed using unpaired Student’s t -tests. * P < 0.05; ** P < 0.01; TGF-β1/MSCs group vs control group.

Article Snippet: Fluorescence-labeled monoclonal antibodies anti-CD4, anti-CD25, anti-CD34, anti-CD105, anti-CD44, anti-CD29, and anti-CD90 were, respectively, added and incubated in the refrigerator at 4°C for 30 min. All antibodies for flow cytometry were purchased from BD Biosciences.

Techniques: Cell Culture, Enzyme-linked Immunosorbent Assay, Expressing, Western Blot, Reverse Transcription Polymerase Chain Reaction

Journal: American Journal of Physiology - Heart and Circulatory Physiology

Article Title: Elevated bone marrow sympathetic drive precedes systemic inflammation in angiotensin II hypertension

doi: 10.1152/ajpheart.00510.2018

Figure Lengend Snippet: Antibodies used for flow cytometry and brain immunohistochemistry analysis in rat

Article Snippet: 9 , Mouse anti-rat CD25:RPE , FC , Bio-Rad , mca273pe.

Techniques: Flow Cytometry, Immunohistochemistry

Journal: Stem Cell Research & Therapy

Article Title: Multiple intravenous injections of allogeneic equine mesenchymal stem cells do not induce a systemic inflammatory response but do alter lymphocyte subsets in healthy horses

doi: 10.1186/s13287-015-0050-0

Figure Lengend Snippet: Multiple allogeneic mesenchymal stem cell (MSC) injections result in changes in splenic regulatory T cell percentages. (A-D) There were no significant changes in splenic CD21 + B-cell (A) , CD4 + T-cell (B) , or CD8 + T-cell percentages (C) or CD4/CD8 ratios (D) following multiple MSC injections. (E) There were no significant changes in activated (CD25 + ) lymphocyte proportions. (F) There were significantly higher percentages of splenic FoxP3 + regulatory T cells in the horses injected with bone marrow (BM)-derived MSCs compared with horses injected with adipose tissue (AT)-derived MSCs. Data are presented as mean ± standard error of the mean. * P <0.05.

Article Snippet: The following antibodies were used: mouse-anti-equine CD3 (clone UC F6G 1:250; Jeffery Stott, University of California, Davis, CA, USA) [ ], mouse-anti-human CD21 (clone B-ly4 1:20; BD Pharmingen, San Jose, CA, USA) [ , ], polyclonal goat-anti-human CD25 (clone AF-223; R&D Systems) [ ], rat-anti-mouse/human FoxP3 (clone PCH101; ebioscience, San Diego, CA, USA) [ ], mouse-anti-CD4 (clone HB61A 1:133; VMRD, Pullman, WA, USA) [ ], mouse-anti-equine CD8 (clone F18P 1:500; J. Stott) [ ], and a donkey-anti-mouse secondary when necessary (1:50; Jackson ImmunoResearch Laboratories, Inc., West Grove, PA, USA).

Techniques: Injection, Derivative Assay