Home › Cell Biology › How to Freeze Cells: Cell Culture Basics Training
Steps
  1. 1 Label vials with cell information 00:06
  2. 2 Prepare freezing materials and equipment 00:25
  3. 3 Remove and wash adherent cells 00:42
  4. 4 Dissociate and detach cells with TrypLE 01:07
  5. 5 Collect cells and prepare suspension 01:33
  6. 6 Count cells for viability assessment 01:56
Cell Biology Thermo Fisher Scientific

How to Freeze Cells: Cell Culture Basics Training

Protocol
Difficulty
intermediate

Steps

1
Label vials with cell information

Label each freezing vial with the cell type, passage number, and date using a permanent marker. This ensures proper identification and tracking of your frozen cell stock.

▶ 00:06
2
Prepare freezing materials and equipment

Gather pre-chilled freezing medium and a Mr. Frosty container filled with isopropanol. These materials are essential for the controlled-rate freezing process.

▶ 00:25
3
Remove and wash adherent cells

Aspirate and discard the conditioned media from culture plates, then wash the attached cells with PBS to remove residual medium.

▶ 00:42
4
Dissociate and detach cells with TrypLE

Add TrypLE to dissociate the culture and detach cells from the flask. Tap the flask gently if needed to dislodge cell clumps.

▶ 01:07
5
Collect cells and prepare suspension

Transfer cells from multiple plates into one or two 15 milliliter centrifuge tubes. Add medium to the cells and mix gently.

▶ 01:33
6
Count cells for viability assessment

Take a small sample of the cell suspension for cell counting to determine cell concentration and viability before freezing.

▶ 01:56
💬 Comments coming soon