How best should I thaw PromoCell hMNC? Can you please send me a protocol?
The following protocol is recommended for the thawing and recovery of PromoCell human Mononuclear Cells (hMNC):
- Refer to the cell number stated in the lot-specific Certificate of Analysis (CoA) and calculate the volume of medium required to achieve a final cell concentration of 1×10⁶ hMNC/ml.
- Transfer the calculated volume of culture medium (e.g. Mononuclear Cell Medium, C‑28030) into a suitable culture vessel and equilibrate for 30 minutes at 37°C and 5% CO₂.
- Remove the cryovial from liquid nitrogen storage and immediately place it on dry ice, even during short transport.
- Thaw the vial in a 37°C water bath for 2 minutes.
- Disinfect the outside of the vial with 70% ethanol and open it in a laminar flow cabinet.
- Using a 2 ml serological pipette, transfer the cell suspension into the pre-warmed medium without resuspending the cells.
- Return the culture vessel to the incubator and culture the cells at 37°C and 5% CO₂.
- Do not resuspend the cells during the thawing process, as this may promote cell clumping.
- For optimal recovery, leave the cells undisturbed for at least 18 hours after seeding.
- Perform the first medium change 18-24 hours after thawing.
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