Natural Killer Cell Isolation from Liver Tissue for Selective Expansion

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. 

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%