mab against human cd25  (Miltenyi Biotec)

Journal: Stem Cell Research & Therapy

Article Title: Towards an advanced therapy medicinal product based on mesenchymal stromal cells isolated from the umbilical cord tissue: quality and safety data

doi: 10.1186/scrt398

Figure Lengend Snippet: Immunosuppression of UCX® and UCX®-ATMP. (A) Suppression of T cell activation was assayed by incubating CD3, CD28 and IL2 activated peripheral blood mononucleated cells (PBMCs) from two different donors with irradiated UCX® and UCX®- advanced therapy medicinal product (ATMP) and measuring lymphocyte proliferation (B) Treg conversion was measured in the same cells by culturing irradiated cells with CD3 + CD4 + CD25 - T-cells and comparing the percentage of converted CD3 + CD4 + CD25 + Foxp3 + regulatory T-cells with unstimulated cells (negative control) or induced conversion with TGFβ. Results are represented as mean ± SEM.

Article Snippet: After five days in culture at 37°C with 5% CO 2 , cells were stained with mAbs against human CD3, CD4 and CD25 (eBioscience) and then stained for huFoxp3 as described by the manufacturer (eBioscience).

Techniques: Activation Assay, Irradiation, Negative Control

Journal: Stem Cells International

Article Title: What Makes Umbilical Cord Tissue-Derived Mesenchymal Stromal Cells Superior Immunomodulators When Compared to Bone Marrow Derived Mesenchymal Stromal Cells?

doi: 10.1155/2015/583984

Figure Lengend Snippet: UCX are more potent immunosuppressors than BM-MSCs. (a) Activation of PBMCs from two different donors was induced by incubation with anti-CD3, anti-CD28, and IL-2 (control). Suppression of lymphocyte proliferation was analysed in the presence of irradiated non-MSCs (Molt4), BM-MSCs, and UCX (both naïve and primed with 10 ng/mL of TNF α and IFN γ ). Independent of donor, lymphocyte proliferation was significantly suppressed when cells were cocultured with MSCs, but not with non-MSCs. Values are represented as mean ± s.e.m. and statistically significant differences are indicated by asterisks (nonparametric t -test Mann Whitney, ∗ P < 0.05). (b) Treg conversion was assayed by using FACS-sorted CD3 + CD4 + CD25 − T cells that were activated with anti-CD3, anti-CD28, and IL-2 alone and with the addition of TGF- β . The effect of irradiated UCX and BM-MSCs in T cell conversion was analysed in cultures without exogenous TGF- β by flow cytometry analysis of CD25 and Foxp3 expression. Values are represented as mean ± s.e.m. and statistically significant differences are indicated by asterisks (unpaired t -test, ∗ P < 0.05).

Article Snippet: After 5 days in culture at 37°C with 5% CO 2 , cells were stained with mAbs against human CD3, CD4, and CD25 (eBioscience, San Diego, CA, USA) and then stained for huFoxp3 as described by the manufacturer (eBioscience, San Diego, CA, USA).

Techniques: Activation Assay, Incubation, Control, Irradiation, MANN-WHITNEY, Flow Cytometry, Expressing

Journal: Stem Cells International

Article Title: What Makes Umbilical Cord Tissue-Derived Mesenchymal Stromal Cells Superior Immunomodulators When Compared to Bone Marrow Derived Mesenchymal Stromal Cells?

doi: 10.1155/2015/583984

Figure Lengend Snippet: UCX are more potent immunosuppressors than BM-MSCs. (a) Activation of PBMCs from two different donors was induced by incubation with anti-CD3, anti-CD28, and IL-2 (control). Suppression of lymphocyte proliferation was analysed in the presence of irradiated non-MSCs (Molt4), BM-MSCs, and UCX (both naïve and primed with 10 ng/mL of TNF α and IFN γ ). Independent of donor, lymphocyte proliferation was significantly suppressed when cells were cocultured with MSCs, but not with non-MSCs. Values are represented as mean ± s.e.m. and statistically significant differences are indicated by asterisks (nonparametric t -test Mann Whitney, ∗ P < 0.05). (b) Treg conversion was assayed by using FACS-sorted CD3 + CD4 + CD25 − T cells that were activated with anti-CD3, anti-CD28, and IL-2 alone and with the addition of TGF- β . The effect of irradiated UCX and BM-MSCs in T cell conversion was analysed in cultures without exogenous TGF- β by flow cytometry analysis of CD25 and Foxp3 expression. Values are represented as mean ± s.e.m. and statistically significant differences are indicated by asterisks (unpaired t -test, ∗ P < 0.05).

Article Snippet: For the induction of Tregs, PBMCs were collected from the Ficoll gradient after centrifugation at 720 g for 30′ at RT, washed with PBS containing 2% FCS, and then stained with mAbs against human CD3, CD4, and CD25 (eBioscience, San Diego, CA, USA) for cell sorting.

Techniques: Activation Assay, Incubation, Control, Irradiation, MANN-WHITNEY, Flow Cytometry, Expressing

Journal: Stem Cell Research & Therapy

Article Title: Towards an advanced therapy medicinal product based on mesenchymal stromal cells isolated from the umbilical cord tissue: quality and safety data

doi: 10.1186/scrt398

Figure Lengend Snippet: Immunosuppression of UCX® and UCX®-ATMP. (A) Suppression of T cell activation was assayed by incubating CD3, CD28 and IL2 activated peripheral blood mononucleated cells (PBMCs) from two different donors with irradiated UCX® and UCX®- advanced therapy medicinal product (ATMP) and measuring lymphocyte proliferation (B) Treg conversion was measured in the same cells by culturing irradiated cells with CD3 + CD4 + CD25 - T-cells and comparing the percentage of converted CD3 + CD4 + CD25 + Foxp3 + regulatory T-cells with unstimulated cells (negative control) or induced conversion with TGFβ. Results are represented as mean ± SEM.

Article Snippet: After five days in culture at 37°C with 5% CO 2 , cells were stained with mAbs against human CD3, CD4 and CD25 (eBioscience) and then stained for huFoxp3 as described by the manufacturer (eBioscience).

Techniques: Activation Assay, Irradiation, Negative Control