Isolation of Murine Lymphocytes

Single-cell suspensions of spleen and lymph nodes were prepared as previously described ^{9 (link), 10 (link)} and resuspended in R10 medium (RPMI 1640 with 10% FBS, 2 mM L-glutamine, 100 U/ml penicillin, 100 μg/ml streptomycin, 10 mM HEPES, 1 mM sodium pyruvate, and 50 μM β-mercaptoethanol). Colon lamina propria lymphocytes were prepared using the Lamina Propria Dissociation Kit (Miltenyi Biotec, Auburn, CA) according to the manufacturer’s instructions. Cells were then isolated using a 40/75% discontinuous Percoll gradient, washed, and resuspended in R10 medium.

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Yang W., Gibson S.A., Yan Z., Wei H., Tao J., Sha B., Qin H, & Benveniste E.N. (2020). Protein Kinase 2 (CK2) Controls CD4+ T-cell Effector Function in the Pathogenesis of Colitis. Mucosal immunology, 13(5), 788-798.

Publication 2020

Lamina Propria Dissociation Kit

Cells Colon Glutamine Hepes Lamina propria Lymph nodes Lymphocytes Mercaptoethanol Penicillin Percoll Pyruvate Sodium Spleen Streptomycin

Corresponding Organization :

Other organizations : University of Alabama at Birmingham, University of North Carolina at Chapel Hill

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Variable analysis

independent variables

Preparation of single-cell suspensions of spleen and lymph nodes
Preparation of colon lamina propria lymphocytes using the Lamina Propria Dissociation Kit

dependent variables

Not explicitly mentioned

control variables

Use of R10 medium (RPMI 1640 with 10% FBS, 2 mM L-glutamine, 100 U/ml penicillin, 100 μg/ml streptomycin, 10 mM HEPES, 1 mM sodium pyruvate, and 50 μM β-mercaptoethanol) to resuspend cells
Isolation of cells using a 40/75% discontinuous Percoll gradient

controls

Positive control: Not specified
Negative control: Not specified

Annotations

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