Isolation and Analysis of Pancreatic Islet Cells

Pancreatic islet tumors of RIP-Tag2 mice perfused with PBS after anesthesia and were removed, weighed, and digested in collagenase II and IV solution (625U/ml, Gibco) with DNase (60 U/ml, Roche), followed by erythrocyte lysis (24 (link)). Dissociated cells were counted and stained for flow cytometric assessment of CD4+, CD8+, Foxp3+, and NK1.1+ cells by FACS (BD LSR Fortessa™) and analyzed with FlowJo software (8.8.6 and X) (details in Supplementary Methods).

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Inoue T., Byrne T., Inoue M., Tait M.E., Wall P., Wang A., Dermyer M.R., Laklai H., Binder J.J., Lees C., Hollingsworth R., Maruri-Avidal L., Kirn D.H, & McDonald D.M. (2021). Oncolytic Vaccinia Virus Gene Modification and Cytokine Expression Effects on Tumor Infection, Immune Response, and Killing. Molecular Cancer Therapeutics, 20(8), 1481-1494.

Publication 2021

collagenase II and IV solution DNase

Anesthesia Cells Collagenase Dnase Erythrocyte Flow cytometric Mice Pancreatic islet tumors

Corresponding Organization :

Other organizations : University of California, San Francisco, UCSF Helen Diller Family Comprehensive Cancer Center, Pfizer (United States)

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Variable analysis

independent variables

Perfusion with PBS after anesthesia
Digestion of islet tumors in collagenase II and IV solution (625U/ml) with DNase (60 U/ml)
Erythrocyte lysis

dependent variables

Tumor weight
Proportions of CD4+, CD8+, Foxp3+, and NK1.1+ cells in dissociated tumor cells measured by flow cytometry

control variables

RIP-Tag2 mouse model
Flow cytometry using BD LSR Fortessa™ and FlowJo software (8.8.6 and X)

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