Catalog Number: AS-15

AMMS^® CIK Cell Expansion Reagent(Product No:AS-08）

AMMS^® CIK Serum-free Medium(Product No:AS15-2)

Product use

Culturing process:

Notes：

* Preparation of amplification solution: before the first feeding (day4), add one recombinant human IL-2 protein (high efficiency type) to the opened first bottle of culture medium after redissolving.

* Feeding time: there is no abnormality observed under the microscope, the cells grow well, the medium turns yellow, and can start to amplification. If the growth is average or poor, consider reducing the amplification or delaying the amplification.

Seeding CIK primary cells (day 0):

1. Collect blood and separation PBMC, inactivate plasma for future use.

2. Seed the cells (Seed density 1x10⁶ cells/ml) into the culture flask with serum-free medium, with a final volume of 30~50ml. Add CIK reagent A and 1.5~5ml of inactivated plasma (5% of the final volume).

3. Put the culture bottle into the CO₂ incubator for culture.

CIK cell activation induced amplification:

Day1: Add CIK reagent B, put into CO₂ incubator for further culture.

Day4: Feed about 100mL, add inactivated plasma 5~10mL. It can also be replenished according to the density. The density is between 0.5~ 1× 10⁶ cells/mL after feeding. (Total volume 150mL).

Day6: Add inactivated plasma about 10~20ml，evenly transfer the cell suspension to the culture bags and feeding. (Total volume 450mL).

Day8: Feed 550mL.Bacteria detection. (Total volume 1000mL).

Day11: Divide the cell suspension evenly into two culture bags and feed equal volume. (Total volume 2000mL).

Day13: Bacteria, endotoxin and mycoplasma detection.

Day14/15: Harvest cells.