Cell Freezing Protocol
Proper cell freezing techniques play a fundamental role in preserving cell viability, maintaining genetic integrity, enabling long-term storage and distribution, and enhancing the efficiency and reproducibility of experiments in biological and medical research.
Proper cell freezing techniques play a fundamental role in preserving cell viability, maintaining genetic integrity, enabling long-term storage and distribution, and enhancing the efficiency and reproducibility of experiments in biological and medical research.
Background
Cryopreservation is a cornerstone of cell biology and is essential to maintaining healthy and reliable cell lines. Freezing cells provides several benefits including: reducing the need for continuous culture maintenance, which can be costly and laborious, facilitating transportation and distribution of cells, enabling long-term storage, and minimizing variability in cell lines due to repeated passaging. BPS Bioscience Cell Freezing Medium is optimized for maximum viability and rapid recovery of all BPS Bioscience cell lines.
Protocol
Tips
- For best results, it is highly recommended to use validated and optimized media from BPS Bioscience. Other preparations or formulations of media may result in suboptimal performance.
- When receiving a new cell line, it is recommended to expand the cells and freeze at least 10 vials at an early passage for future use.
Adherent Cell Freezing Protocol
This protocol is applicable to adherent cell lines, such as HEK293, CHO or HeLa cells.
- Aspirate the medium, wash the cells with phosphate buffered saline (PBS) without Ca2+ and Mg2+, and detach the cells from the culture vessel with 0.05% Trypsin/EDTA or an alternative detachment solution.
- Once the cells have detached, add growth medium and count the cells.
- Spin down the cells at 300 x g for 5 minutes, remove the medium and resuspend the cells in 4°C Cell Freezing Medium (#79796) at ~2 x 106 cells/ml.
- Dispense 1 ml of cell aliquots into cryogenic vials. Place the vials in an insulated container for slow cooling and store at -80°C overnight.
- Transfer the vials to liquid nitrogen the next day for storage.
Suspension Cell Freezing Protocol
This protocol is applicable to suspension cell lines, such as Jurkat, THP-1, etc. or PBMCs.
- Spin down the cells at 300 x g for 5 minutes, remove the medium and resuspend the cell pellet in 4°C Cell Freezing Medium (#79796) at a density of ~2 x 106 cells/ml.
- Dispense 1 ml of cell aliquots into cryogenic vials. Place the vials in an insulated container for slow cooling and store at -80°C overnight.
- Transfer the vials to liquid nitrogen the next day for storage.
