2024.02.18
Please optimize the process by following these steps:
• Follow the User Guide Step by Step: The first and foremost step to improve thawed cell viability is to adhere strictly to the thawing protocol provided by the cell supplier or outlined in the user guide.
• Ensure Rapid Thawing at 37°C: Rapidly thaw the cells in a 37°C water bath to minimize the time cells spend in the presence of damaging ice crystals and to reduce the osmotic shock experienced during thawing. This step should take no longer than necessary to just melt the ice, typically 1-2 minutes.
• Gentle Handling to Minimize Mechanical Stress: After thawing, gently mix the cells with pre-warmed culture medium to dilute the cryoprotectant gradually and avoid osmotic shock. Avoid vigorous pipetting or shaking, as mechanical stress can significantly reduce cell viability.
• Centrifuge Cells Under Optimal Conditions: Gently dilute the thawed cell suspension with warm culture medium. Then, centrifuge the cell suspension following the specific guidelines recommended for the cell type, using mild conditions to pellet the cells and remove the supernatant containing the cryoprotectant. Exercise caution regarding centrifuge speeds and durations to prevent mechanical damage to the cells.
• Carefully Resuspend and Culture Cells: After discarding the supernatant, carefully resuspend the cell pellet in a fresh, pre-warmed culture medium. Use gentle pipetting to avoid creating shear stress that can damage the cells. Transfer the cells to a suitable culture vessel and store or incubate under optimal conditions for the specific cell type.
• Option: Use an optimal, such as Percoll-Containing Thawing Medium: Incorporating a Percoll-containing medium as suggested for thawing and purification can significantly enhance cell viability. This approach not only aids in gentle separation and recovery of viable cells but also minimizes the exposure to detrimental conditions during the thawing process.
