Neutrophils require a different approach to cell thawing and handling because they are so easily activated and damaged. Follow the procedure below to ensure high post-thaw viability.
Materials
- Vial of frozen neutrophils
- 37°C water bath
- Hank’s Balanced Salt Solution (HBSS)
- Centrifuge
- Tubes, pipets
Procedure
- Place vial of cells in 37°C water bath and gently swirl until thawed (approximately 2 minutes). It is important to thaw the cells quickly. Do NOT allow thawed cells to remain in freezing media any longer than necessary.
- In a 15 mL conical tube, slowly add freshly thawed cells to 9 mL of HBSS. DO NOT USE MEDIUM-CONTAINING SERUM. Invert tube 2 or 3 times to mix, or mix gently by pipetting up and down several times.
- Centrifuge for 10 minutes at 200 x g.
- Aspirate or decant the supernatant and gently resuspend the cell pellet in 10 mL of HBSS.
- Remove an aliquot for cell count and proceed with experimental manipulations.
- Neutrophils can be activated by endotoxin, excessive agitation, or repeated centrifugation with cell death resulting. Use of polypropylene tubes is recommended. Minimize centrifugation steps and use promptly after thawing.
Related Resources
Interested in learning more protocols and research tips? Then check out our related content on thawing cells and performing other common laboratory procedures.
Recent Posts
Site Search
Tag Cloud
Assays
B Cells
Cardiovascular Disease
Cell Biology
Cell Isolation
Cell Therapy
Cryopreserved
Dendritic Cells
Drug Discovery
eBook
Fresh Cells
Gene Therapy
HLA System
Immunology
Monocytes
NK Cells
PBMC
Pooled PBMCs
Quality
Regulations
Regulatory T Cells
Research Ethics
T cells
TIL Therapy
Tips & Tricks
Transplantation