KBM NK (Complete medium and kit for culturing NK cells selectively from human PBMCs)

Cosmo Bio LTD

Catalog No: KJN-16030210

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Overview
KBM NK is the complete medium and kit for culturing NK cells selectively from human PBMCs (Peripheral Blood Mononuclear Cells) derived from human peripheral blood. It includes KBM NKCC-b (culture bag), KBM NKCC-c (coating agent), KBM NKCC -1 (Medium for primary culture), and KBM NKCC -2 (Medium for expansion culture.

Description
KBM NK kit is a kit that selectively cultivates NK cells from PBMC (Peripheral Blood Mononuclear Cells) derived from human peripheral blood efficiently and easily. The kit is conposed by; KBM NKCC-b (culture bag)
KBM NKCC-c (coating agent)
KBM NKCC -1 (Medium for primary culture)
KBM NKCC -2 (Medium for expansion culture)
Refer to the operation manual (Link) for the proper use.

Product Features
In 10 days of culture, it is possible to multiply cells by 100 to 150 times (Up to 3.6 billion cells at maximum), and more than 50% of them become NK cells (*).
* Rarely, the NK ratio does not exceed 50% due to individual differences.
 
Method of Usage
Example of Culture Protocol
1. PBMC 2.0E+07 cells were seeded into T75 flasks pre-coated (NKCC-1 20mL, serum 2mL added) and incubated in a 5% CO2 incubator at 37℃.
2. After 5 days of culture, the cells were collected when the number of cells reachs to 8.2E+07 cells. Transfer to expansion culture (1200 rpm x 5 min).
3. Add 200 mL of NKCC-2 so that the cell concentration gets 4.0E+05 cells/mL. Add inactivated (56℃, 30 min) human serum (10 mL) obtained at the time of PBMC separation.
4. After 7 days of culture, add the remaining 800nL of NKCC-2 to the bag, after it was confirmed that the medium had become yellowish.
5. After 10 days of culture, count the number of cells, and confirm the cell surface markers (CD56, CD3) and measure the cytotoxic activity using K 562 (target cancer cell) on a flow cytometer.