AMMS®NK Cell Culture Kit 3.0

Natural Killer (NK) cell therapy has become a pivotal research focus in immunotherapy, demonstrating remarkable potential in oncology. NK cells are innate lymphocytes with broad-spectrum cytotoxic activity, capable of targeting tumor cells, virus-infected cells, and other pathological entities. Distinct from conventional T cell therapies, NK cells exert immediate cytotoxic effects without prior antigen priming, enabling rapid therapeutic action independent of tumor antigen specificity.

Figure 1. Anti-tumor Mechanisms of NK Cells (Image source: Acta Pharmaceutica Sinica)

NK cells can be sourced from diverse origins, including peripheral blood, umbilical cord blood (UCB), induced pluripotent stem cells (iPSCs), and the NK92 cell line. Studies highlight that UCB-derived NK cells and progenitors exhibit the highest abundance among available sources. The naïve phenotype of UCB-derived NK cells reduces Graft-versus-Host Disease (GVHD) risks, while their broad availability and superior proliferative capacity position UCB as a strategic source for NK cell production. Notably, the first CAR-NK clinical trial employed UCB-derived NK cells. The growing prominence of UCB-NK cells in immunotherapy research underscores the critical challenge of achieving scalable, high-purity NK cell expansion in vitro for clinical translation.

The AMMS® NK Cell Culture Kit 3.0 is designed for ex vivo activation and expansion of high-purity NK cells from fresh or cryopreserved umbilical cord blood mononuclear cells (CBMCs). A 2-liter culture system achieves yields of 4-6 billion cells with NK purity exceeding 90%.

Optimised for Fresh/ Frozen Cord Blood, CBMC, Frozen PBMC

AMMS® NK Cell Culture Kit 3.0-A/D: −20 °C

AMMS® NK Cell Culture Kit 3.0-E/G: 2–8 °C

AMMS® NK Activation Culture Medium ×1: 2–8 °C, protect from light

AMMS® NK Expansion Culture Medium ×2: 2–8 °C, protect from light

1. Sample requirements:

   ① If the blood volume of umbilical cord blood (including anticoagulant) is less than 80mL or the expected NK harvest is more, it is recommended to use HPL instead of autologous plasma. The cell proliferation rate of HPL is about 1-2 times higher than that of autologous plasma.

   ② The operation should be completed within 12 hours after blood collection.

   ③ It is recommended that the freezing density of frozen samples be 2-4 x 10 7/mL, and the viability rate after recovery should not be lower than 80%. HPL should be used instead of autologous plasma during the cultivation of umbilical cord blood samples. Autologous plasma should be used for peripheral blood samples (heparin sodium anticoagulant), and HPL can also be used instead of autologous plasma.

2. Vessel density: the initial cell density of the bottle is recommended to be 1x10 6 cells/mL. When there are more red blood cells, the fluorescence counting or red blood cell lysis counting should be selected to avoid affecting the bottle density of the cell.

3. Fluid density: the density after fluid replacement should not be lower than 1x10 6/mL.

4. Use of culture medium:

   ① The culture medium should be naturally rewarmed at room temperature before each infusion.

   ② Do not put the whole bottle of culture medium into the 37℃ incubator for re-temperature, otherwise it will accelerate the inactivation of cytokines in the supplemented culture medium.

   GMP Grade CGT Reagent

   ③ The prepared complete medium has a short shelf life, and it is recommended to use it up in about one week.

5. Proper handling and preservation of plasma: see instructions for details. Centrifuged plasma should be clear.

6. Use of culture bags: When the culture volume is less than 1L, the culture bags need to be folded before being placed. It is recommended to use the recommended model of our company.

7. Incubation time: A, the factor should be incubated for 4 ℃ overnight in a flat bed. (In case of emergency, try to incubate 37℃ for 2 hours)

8. Do not shake the culture bottle at will during the initial stage of culture: otherwise, the activated clone group is easy to float up, and the coating factor will reduce the activation of the cell group.

9. Use of factors: In order to reduce the loss of factors hanging on the wall, it is recommended to perform centrifugation before use. Place the vial containing factors into a 50mL centrifuge tube and centrifuge at 1000rpm for 1-2min.

10. Equipment maintenance: check the temperature and concentration of CO2 incubator regularly, and replace the filter in time. Regular maintenance and cleaning of biological safety cabinet.

11. Environmental monitoring: replace primary, medium and high efficiency filters regularly to ensure the environmental standards of clean areas.

12. Fixed types and models of experimental consumables: the impact of changing models and specifications on cultivation effects should be evaluated in advance, such as 75cm2 culture bottles and cell culture bags.