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Nexcelom cellometer auto t4

Manufactured by Revvity

The Nexcelom Cellometer Auto T4 is a cell counting and analysis instrument. It utilizes automated cell counting and analysis technology to determine the concentration and viability of cells in a sample.

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3 protocols using nexcelom cellometer auto t4

1

Adoptive Transfer of OT-I T Cells

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OT-I Thy1.1+ TCR transgenic T cells were harvested from spleen and mLN of naïve animals. Flow cytometry was used to determine the frequency of OT-I T cells prior to adoptive transfer by staining with anti-Vα2 (used by both TCRs) and anti-CD8 (BD Pharmingen, San Diego, CA). Cells were counted with a Nexcelom Cellometer Auto T4 (Nexcelom Bioscience, Lawrence, MA), and 106 WT OT-I T cells with our without the same number of OT-II T cells were transferred into naive C57BL/6 mice i.v. 24 to 48 hours prior to infection or transplantation.
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2

PDGF-BB and SDF-1α Mediated Cardiac Fibroblast Proliferation

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Cells were maintained in DMEM/F12 containing 10% fetal bovine serum under standard tissue culture conditions. Subconfluent cultures in 24-well plates were growth-arrested for 2 days in DMEM/F12 containing 0.4% bovine serum albumin. Next cells were placed in DMEM/F12 containing a low concentration (25 ng/mL) of PDGF-BB and then treated every day for 4 days without or with various treatments. In experiments in which cells were treated with SDF-1α, the SDF-1α was co-administered with sitagliptin (1 μmol/L). Sitagliptin was administered with SDF-1α because we have found that sitagliptin, by blocking DPP4, prevents the metabolic inactivation of SDF-1α and thus enhances its effects on proliferation of cardiac fibroblasts.13 (link) Finally, cells were harvested and cell number quantified using a Nexcelom Cellometer Auto T4 cell counter (Nexcelom Bioscience: Lawrence, MA).
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3

Adoptive Transfer of OVA-specific T Cells

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To monitor antigen-specific CD8+ T cell responses, we adoptively transferred OVA-specific transgenic T cells into naive prior to inoculation with B16 melanoma expressing OVA. For the adoptive transfer of tumor-specific CD8+ T cells, spleens and mesenteric lymph nodes from CD45.2+Thy1.2+Fcgr2b–/– OT-I mice or CD45.2+Thy1.1+ WT OT-I mice were processed into single-cell suspensions. Cells were counted using a Nexcelom Cellometer Auto T4 (Nexcelom Bioscience) and stained with CD8-BV785, CD4-PacBlue, Thy1.1-PerCP, CD45.2-PE/Dazzle, CD45.1-BV605, Vα2-FITC, and Vβ5-PE (all from BioLegend). The frequency of OT-I and OT-II cells was determined via Vα2 and Vβ5 TCR coexpression. Cells were then resuspended in 1× PBS and 1 × 106 OT-I and 1x106 OT-II cells were transferred intravenously into naive congenic CD45.1+ B6-Ly5.1/Cr hosts 24 hours prior to tumor inoculation.
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