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).