Quantifying CD4+ and CD8+ T-cell Populations

The analysis was carried out as described previously [11 (link)]. In brief, the enzymatically digested (collagenase, hyaluronidase type IV-S and DNase I) pancreatic tissue samples (n = 3) were mashed through nylon mesh cell strainers (70 µm). Single-cell sample suspensions devoid of red blood cells were treated for 20 min/RT with antibodies, fluorescently labeled and analyzed (no less than 4 × 10⁴ cells) using flow cytometry (BD FACS Canto II) to quantify the percentage of CD4+ and CD8+ populations in pancreas and blood specimens. Antibodies (BioLegend, San Diego, CA, USA) were used according to the manufacturer’s instructions: PerCP/Cyanine5.5 anti-mouse CD45 (clone 30-F11), phycoerythrin (PE) anti-mouse CD3 (clone 17A2), fluorescein isothiocyanate (FITC) anti-mouse CD4 (clone GK1.5) and APC/Cyanine7 anti-mouse CD8a (clone 53-6.7).

Free full text: Click here

Jazowiecka-Rakus J., Sochanik A., Hadryś A., Fidyk W., Chmielik E., Rahman M.M, & McFadden G. (2022). Combination of LIGHT (TNFSF14)-Armed Myxoma Virus Pre-Loaded into ADSCs and Gemcitabine in the Treatment of Experimental Orthotopic Murine Pancreatic Adenocarcinoma. Cancers, 14(8), 2022.

Publication 2022

BD FACS Canto II PE) anti-mouse CD3 (clone 17A2),

Anti cd3 Antibodies Blood Cells Clone Collagenase Dnase i Flow cytometry Fluorescein Hyaluronidase Isothiocyanate Mouse Nylon Pancreas Phycoerythrin Populations Red blood cells Tissue

Corresponding Organization : The Maria Sklodowska-Curie National Research Institute of Oncology

Other organizations : Arizona State University

Top 5 similar protocols

Variable analysis

independent variables

Enzymatic digestion of pancreatic tissue samples (collagenase, hyaluronidase type IV-S and DNase I)

dependent variables

Percentage of CD4+ and CD8+ populations in pancreas and blood specimens

control variables

Mashing pancreatic tissue samples through nylon mesh cell strainers (70 µm)
Treating single-cell sample suspensions devoid of red blood cells for 20 min/RT with antibodies
Fluorescently labeling the cells
Analyzing no less than 4 × 10^4 cells using flow cytometry (BD FACS Canto II)

Annotations

Based on most similar protocols

Etiam vel ipsum. Morbi facilisis vestibulum nisl. Praesent cursus laoreet felis. Integer adipiscing pretium orci. Nulla facilisi. Quisque posuere bibendum purus. Nulla quam mauris, cursus eget, convallis ac, molestie non, enim. Aliquam congue. Quisque sagittis nonummy sapien. Proin molestie sem vitae urna. Maecenas lorem.

As authors may omit details in methods from publication, our AI will look for missing critical information across the 5 most similar protocols.

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Revolutionizing how scientists
search and build protocols!