Catalog Number: AS-22
AMMS® NK Cell Culture Kit. 2.0(Product No:AS22- 1）
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Component Descriptions |
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| Component Name | Cat. No. | Specification | Amount | Storage | Product characteristics | Shelf Life |
| NK reagent A-2.0 | AS22- 1A | 200μL | 1 stick | -20℃ | Liquid | 18 Months |
| NK reagent B-2.0 | AS22- 1B | 500μL | 1 stick | -20℃ | Liquid | 18 Months |
| NK reagent C-2.0 | AS22- 1C | 500μL | 1 stick | -20℃ | Liquid | 18 Months |
| NK reagent D-2.0 | AS22- 1D | 500μL | 1 stick | -20℃ | Liquid | 18 Months |
AMMS® NK Serum-free Medium(Product No:AS01-2)
| Product name | Cat. No. | Specification | Amount | Storage | Product characteristics | Shelf Life |
| AMMS® NK Serum-free Medium | AS01-2 | 1000mL | 2 flasks | 2~8℃ | Liquid | 18 Months |
Description
This product is suitable for autologous peripheral blood or umbilical cord blood PBMC, through in vitro activation and amplification to obtain higher purity of NK cells. Only for in vitro research use.
Product use
| Steps | Cultivation time | Use of reagents | Cultivation container | Complete culture medium | Inactivated plasma | Total volume | Remarks |
| Coating | Day -1 | NK reagent A-2.0 | 175cm2 Culture flask | / | / | / | Keep coating flask flat at 4℃ overnight. |
| Seeding | Day 0 | NK reagent B-2.0 | 175cm2 Culture flask | 22.5mL | 2.5mL | 25mL | Density of seed:
2× 106 pcs/mL |
|
Culture |
Day 3
(1st feed) |
NK reagent C-2.0 | 175cm2
Culture flask |
46.5mL | 3.5mL | 75mL | Do not disperse cells. Do not touch the cell layer at the bottom of the flask when replenishing the medium. |
| Day 5
(2nd feed) |
NK reagent D-2.0 | 175cm2 Culture flask | About 166.25mL | 8.75mL | 250mL | ||
|
Bagging |
Day7
(3rd feed) |
Complete culture medium | Cell culture bag | About 350mL | Remaining plasma | 600mL | It can also be replenished according to the density, The density is between 0.6~ 1× 106 cell/mL after seed. Pat the culture bag after feeding. Make the cells evenly distributed. |
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Split bag
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Day9
(4th feed) |
Complete culture medium | Cell culture bag | Aad to each bag 300 mL | / | 1200mL | |
| Day 11/12 (5th feed) | Complete culture medium | Cell culture bag | Add to each bag 400 mL | / | 2000mL | ||
| Harvest | 14 day、
15 day |
/ | Cell culture bag | / | / | 2000mL | Harvest cells |
Notes：
* The medium should be allowed at room temperature for more than 1h before each use（Disable the relevant equipment to force rapid rewarming），this is the same for subsequent operations.
* Inactivated plasma is calculated at 5% ~ 10% of the culture system. If there are more anticoagulants, it is recommended that the plasma be increased to 7% ~ 12%.
