BINKIT for NK cells expansion from PBMCs

Product features

 Natural killer (NK) cells can be expanded from human peripheral blood mononuclear cells (PBMCs) without using feeder cells.  NK cells can be expanded several hundred to several thousand-fold by 2 - 3 weeks of culture.

 One kit is sufficient to expand NK cells from 20 - 50 ml of whole blood.

Kit name Catalog No. Amount BINKITⓇ N501-1 1 kit N501-2 2 kits N501-4 4 kits N501-8 8 kits

Kit components

Catalog No. Amount One kit of BINKITⓇ includes: NK Cell Initial Flask (75 cm2) N104 (M) 1 flask NK Cell Initial Medium N115a 45 ml NK Cell Initial Cocktail N115b 1.

9 ml NK Cell Subculture Medium N201 1000 ml

Procedures

Preparing reagents

NK Cell Initial Medium and NK Cell Subculture Medium should be supplemented with 5 % (v/v) of heat-inactivated FBS or autologous plasma.

Preparing peripheral blood mononuclear cells (PBMCs)

Isolate PBMCs from whole human blood by density gradient centrifugation using Ficoll-Paque. Washing NK Cell Initial Flasks Add 10 ml PBS to an NK Cell Initial Flask.

Slant the flask to cover the entire surface with PBS.

Aspirate the liquid completely from the flask. Care should be taken not to scratch the surface of the flask. Repeat the washing process two more times.

Culturing NK cells from PBMCs

Suspend the PBMCs in NK Cell Initial Medium at 1  106 cells/ml. Add 40 l of NK Cell Initial Cocktail to the cell suspension.

Transfer the cell suspension to the pre-washed NK Cell Initial Flask. Incubate under 5 % CO2 at 37 C for 3 days.