Natural Killer Cell Isolation from Liver Tissue for Selective Expansion

Published: July 31, 2023

Abstract

Source: Ma, M., et al. Natural Killer (NK) and CAR-NK Cell Expansion Method using Membrane Bound-IL-21-Modified B Cell Line. J. Vis. Exp. (180), e62336, doi:10.3791/62336 (2022).

This video demonstrates the isolation of lymphocytes, including natural killer cells or NK cells, from human liver tissue for selective expansion. Enzymatic and mechanical digestion, followed by density gradient separation, promotes lymphocyte isolation, including NK cells. The collected lymphocytes can be further used for the selective expansion of NK cells.

Protocol

All procedures involving human participants have been performed in compliance with the institutional, national, and international guidelines for human welfare and have been reviewed by the local institutional review board. 1. NK cell expansion from liver tissues (Day 0), as shown in Figure 1. NOTE: Initial cell number and viability are strongly correlated with the time since organ removal and the initial tissue sample amount. How…

Representative Results

Figure 1: Diagram of NK cell expansion from solid human organ samples. Briefly, the obtained human liver samples are minced into small cubes for mechanical digestion. Dissociated cells are then isolated using PVP-coated silica and Lymphocyte Separation Media. 

Divulgazioni

The authors have nothing to disclose.

Materials

Collagenase IV Sigma C4-22-1G For TILs isolation
Hank's Balanced Salt Solution (HBSS – w/o calcium or magnesium) ThermoFisher 14170120 For TILs isolation
Lymphocyte Separation Media Corning 25-072-CV For TILs isolation
Phosphate-Buffered Saline (PBS) Corning 21-040-CV For transduction
R-10 media
Ingredients: RPMI 1640
VWR 45000-404
R-10 media
Ingredient: L-glutamine
VWR 45000-304 1%
R-10 media
Ingredient: Penicillin Streptomycin
VWR 45000-652 1%
R-10 media
Ingredient: Fetal Bovine Serum (FBS)
Corning 35-010-CV 10%

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Citazione di questo articolo
Natural Killer Cell Isolation from Liver Tissue for Selective Expansion. J. Vis. Exp. (Pending Publication), e21556, doi: (2023).

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