Journal: International Journal of Molecular Sciences

Article Title: Proinflammatory Cytokine Preconditioning Enhances the Therapeutic Potency of Different Types of MSCs in Inflammation

doi: 10.3390/ijms27094090

Figure Lengend Snippet: Cytomix increased the expression of immunomodulatory markers on BM-MSCs, iMSCs WT and iMSCs B2M KO. The gating strategy is shown in . ( A ) Histogram charts of the expression of the immunomodulatory markers on BM-MSCs, iMSCs WT and iMSCs B2M KO. ( B – H ) Cytomix significantly increased the expression levels of HLA-ABC on BM-MSCs and iMSCs WT, HLA-DR on BM-MSCs, CD54 on all types of MSCs, CD200 on iMSCs WT, CD119 and CD120b on BM-MSCs, and CD274 on iMSCs WT and iMSCs B2M KO. Representative results of three independent experiments are shown. Results are shown as mean ± SD ( n = 3 in each group) and analysed by two-way ANOVA with Šídák’s multiple comparisons test or Tukey’s multiple comparisons test. */**/***/**** represent comparisons of naïve vs. Cytomix in each type of MSC; $/$$$/$$$$ represent comparisons of BM-MSCs vs. iMSCs WT vs. iMSCs B2M KO in the naïve group; ##/###/#### represent comparisons of BM-MSCs vs. iMSCs WT vs. iMSCs B2M KO in the Cytomix group with significance levels of p < 0.05/0.01/0.001/0.0001, respectively. Nd represents not detectable.

Article Snippet: The cells were centrifuged at 300× g for 5 min and resuspended in 100 μL FACS buffer containing anti-human HLA-ABC-VioGreen, HLA-DR-VioBlue, CD54-APC, CD274-PE-Vio ® 615, CD119-FITC (Cat# 130-120-436, 130-111-794, 130-121-342, 130-122-811, 130-099-931; Miltenyi Biotec, Bergisch Gladbach, Germany), CD120b-PE and CD200-PE-CY7 antibodies (Cat# 358403, 399805; BioLegend, San Diego, CA, USA).

Techniques: Expressing

Journal: STAR Protocols

Article Title: Chimerism and phenotypic analysis of intraepithelial and lamina propria T cells isolated from human ileal biopsies after intestinal transplantation

doi: 10.1016/j.xpro.2023.102192

Figure Lengend Snippet: Primary surface antibody staining panel: Pt25 multilineage panel

Article Snippet: Mouse monoclonal anti-Human HLA-A2, A28 (clone REA142) FITC , Miltenyi Biotec , Cat#130-099-601; RRID: AB_2652040.

Techniques: Staining

Journal: STAR Protocols

Article Title: Chimerism and phenotypic analysis of intraepithelial and lamina propria T cells isolated from human ileal biopsies after intestinal transplantation

doi: 10.1016/j.xpro.2023.102192

Figure Lengend Snippet: Gating strategy for multilineage chimerism, as well as donor and recipient T cell phenotypes, in post-transplant ileal biopsies, using ITx Pt25 POD241 IEL as an example (A) Representative contour plots depicting gating strategy of HLA-ABC + CD45 + cells, CD3 + T cells, CD3 + γδTCR + T cells, CD3 + γδTCR − CD4 + T cells, CD3 + γδTCR − CD8 + T cells, CD19 + B cells, and CD3 - CD56 + NK cells. (B) Percentages of donor (“D”) and recipient (“R”) multilineage chimerism in Pt25 POD241 IELs. Pt25’s donor is HLA-B12 + . Pt25, the recipient, is HLA-A68 + , which has serological cross-reactivity with HLA-A28. (C) Gating strategy for donor (“D”) and recipient (“R”) CD4 + and CD8 + T cells in Pt25 POD241 IELs. CD45RA vs. CCR7 to categorize naïve T cells (CD45RA + CCR7 + ), central memory T cells (TCMs: CD45RA − CCR7 + ), effector memory T cells (TEMs: CD45RA − CCR7 − ), and terminally-differentiated effector memory T cells (TEMRAs: CD45RA + CCR7 - ), tissue-resident memory T cells (TRMs: CD69 + CD103 +/− CD49a +/ − ), effector T cells (CD28 + NKG2D +/− ).

Article Snippet: Mouse monoclonal anti-Human HLA-A2, A28 (clone REA142) FITC , Miltenyi Biotec , Cat#130-099-601; RRID: AB_2652040.

Techniques:

Journal: STAR Protocols

Article Title: Chimerism and phenotypic analysis of intraepithelial and lamina propria T cells isolated from human ileal biopsies after intestinal transplantation

doi: 10.1016/j.xpro.2023.102192

Figure Lengend Snippet: Primary surface antibody staining panel: Pt25 B-cell panel

Article Snippet: Mouse monoclonal anti-Human HLA-A2, A28 (clone REA142) FITC , Miltenyi Biotec , Cat#130-099-601; RRID: AB_2652040.

Techniques: Staining

Journal: STAR Protocols

Article Title: Chimerism and phenotypic analysis of intraepithelial and lamina propria T cells isolated from human ileal biopsies after intestinal transplantation

doi: 10.1016/j.xpro.2023.102192

Figure Lengend Snippet:

Article Snippet: Mouse monoclonal anti-Human HLA-A2, A28 (clone REA142) FITC , Miltenyi Biotec , Cat#130-099-601; RRID: AB_2652040.

Techniques: Recombinant, Saline, Software, Flow Cytometry, Sterility

Journal: STAR Protocols

Article Title: Chimerism and phenotypic analysis of intraepithelial and lamina propria T cells isolated from human ileal biopsies after intestinal transplantation

doi: 10.1016/j.xpro.2023.102192

Figure Lengend Snippet:

Article Snippet: Mouse monoclonal anti-Human HLA-A2, A28 (clone REA142) FITC , Miltenyi Biotec , Cat#130-099-601; RRID: AB_2652040.

Techniques: Concentration Assay, Staining