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產品編號：N501,N520
BINKIT NK cell expansion kit
  • 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 in 2 to 3 weeks of culture.
  • One kit is sufficient to expand NK cells patients using 20-50 mL each of the whole blood.
  • Research use only.
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BINKIT NK cell expansion kit

Contents
  • NK Cell Initial Medium, Cocktail
  • NK Cell Initial Flask
  • NK Cell Subculture Medium

　　BINKIT specification sheet (PDF file)

Options
  • Culture bag(small, large)
  • Culture bag connection tube
MSDS
Application Data
Procedure overview
Procedures
  • ・Preparing reagentsNK 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)
  • PBMCs should be isolated from the whole blood by density gradient centrifugation using Ficoll-Paque.
  • ・Washing an NK cell Initial Flask
  • 10 ml PBS should be added 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 inner surface of the flask. Repeat the washing process two more times.
  • ・Culturing NK cells from PBMCs
  • PBMCs should be suspended in the NK Cell Initial Medium at 1 x 106 cells/ml. Add 40 μl of NK Cell Initial Cocktail to the 1 ml of the cell suspension. Seed the cells onto the pre-washed NK Cell Initial Flask, and incubate under 5 % CO2 at 37 ℃ for 3 days.
  • ・Changing medium and sub-culturing
  • Transfer floating as well as adherent cells to conical centrifuge tube and centrifuge at 200 x g for 8 minutes. Remove the supernatant and re-suspended the cells at 1 x 106 cells/ml in the NK Cell Subculture Medium supplemented with 5 % (v/v) of heat- inactivated FBS or autologous plasma. The cell suspension is transferred to conventional culture flasks and incubated under 5 % CO2 at 37 ℃. Cells should be sub-cultured every 2 - 3 days by suspending cells in the completed NK Cell Subculture Medium at 0.8 x 106 cells/ml.
  • ・Suggested culturing period: 2 to 3 weeks.
Other supplies required
  • Ficoll-Paque (GE Healthcare, Sweden)
  • Sterile PBS
  • FBS or autologous plasma (It is desirable to have them heat-inactivated at 56 ℃ for 30 minutes.)
  • Sterile conical centrifuge tubes
  • Sterile culture flasks
Precautions
  • NK Cell Initial Flask may carry condensation on the surface, which does not adversely affect the performance of the kit.
    This product contains human plasma albumin.
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