Evaluating T-cell Proliferation and Cytokine Production

Mice splenocytes were obtained as described previously (5 (link)). Both primary skin fibroblast-like cells (10⁴) and SphCs (10⁴) were cocultured for 72 h with 5×10⁵ splenocytes activated with anti-CD3 antibody (1 mg/mL, eBioscience). A CellTrace Proliferation Kit (Thermo Fisher Scientific, USA) was used to evaluate T-CD4⁺ proliferation by staining with anti-CD4-APC. Cell media were harvested at 36 h, and cytokines were analyzed by FACS using a Cytometric Bead Array Mouse Th1/Th2/Th17 Cytokine Kit (BD Biosciences).

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Andreone L., dos Santos A.F., Wailemann R.A., Terra L.F., Gomes V.M., Macedo da Silva J., Rosa-Fernandes L., Sogayar M.C., Palmisano G., Labriola L, & Perone M.J. (2023). Cotransplantation of marginal mass allogeneic islets with 3D culture-derived adult human skin cells improves glycemia in diabetic mice. Brazilian Journal of Medical and Biological Research, 56, e12611.

Publication 2023

anti-CD3 antibody CellTrace Proliferation Kit Cytometric Bead Array Mouse Th1/Th2/Th17 Cytokine Kit

Anti antibody Cell Cytokine Fibroblast Mouse Skin Th17

Corresponding Organization :

Other organizations : Austral University, Consejo Nacional de Investigaciones Científicas y Técnicas, Universidade de São Paulo

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Variable analysis

independent variables

Primary skin fibroblast-like cells (10^4)
SphCs (10^4)
Anti-CD3 antibody (1 mg/mL, eBioscience)

dependent variables

T-CD4+ proliferation
Cytokine levels

control variables

Splenocytes (5x10^5)

controls

Positive control: Splenocytes activated with anti-CD3 antibody
Negative control: Not mentioned

Annotations

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