Nunc Cell Factory

Summary

Prepare a cell suspension from the culture solution by injecting the suspension into each of the small chambers of the unit. Place with the long side down. Rotate the cell 90 degrees, flatten and inflate with CO2. Then close and incubate.

Operation method

Nunc Cell Factory

Principle

Prepare a cell suspension from the culture solution by injecting the suspension into each of the small chambers of the unit. Place with the long side facing down. Rotate the cell 90 degrees, flatten it and inflate it with CO2. Then close and incubate.

Materials and Instruments

Monolayer cells 0.25% natural trypsin D-PBSA
Growth medium
Nunc CellWorks Silicone tubing and connectors Hematocrit plate or electronic cell counter and liquid for counting

Move

1. After digestion with trypsin, suspend the cells, count the cells, and dilute the suspension with 2 L of culture medium to a cell density of ^2x104 cells/ml.

2. Place the chambers so that the long side is facing down, connect the culture medium supply tube to the Luer connection tube, and then through the supply tube to the bottom hole.



3. Add cells and culture medium through the supply tube. Liquid will be at the same level in all culture chambers.

4. Rotate the chambers 90 degrees in the plane of the cell monolayer so that the short side of the unit is facing down, and place so that the inlet holes and the supply tubes are facing up.

5. At the Luer Connection Tube, interrupt the connection of the supply tubes to the culture fluid reservoir.

6. Perpendicular to the plane of the cell monolayer, rotate the chambers 90 degrees in the plane of the cell monolayer to level the bottom, so that the culture surfaces are in a horizontal position.

7. Aerate the unit with 5% CO2 air for 5 minutes. _CO2 air to inflate the cell for 5 min, then clamp the supply tube and outlet. If necessary, inflate continuously.

8. Empty the culture solution from the supply tube and outlet, and transfer the unit into the incubator.

9. When replacing the culture solution (or collecting the culture solution), follow the reverse procedure of step 6. Remove the filter on the input tube, and then follow the opposite procedure of step 4.

10. Scrub the Luer connection tube and loosen the clamp to allow the culture solution to flow out.

11. Replace the culture solution according to steps 2~8.

12. Collect the cells.

(a) Remove the culture medium according to steps 9 and 10.

(b) Add 500 ml D-PBSA and remove.

(c) Add 500 ml trypsin at 4°C and remove after 30 s.

(d) Digest the cells with the remaining trypsin for 15 min.

(e) Add culture medium and shake the culture chamber to suspend the cells.

(f) Remove the cell-containing culture medium according to steps 9 and 10.

13. The residual cells can be used to inoculate the next batch of cells, although it is difficult to control the density of inoculation with this method. It is best to discard the culture chambers and restart the culture with new chambers.

For more product details, please visit Aladdin Scientific website.

https://www.aladdinsci.com/