BINKIT (NK cell expansion kit) NK細胞培養資材・試薬
NK細胞培養 NK細胞放大 擴大 擴增 培養基
(We manufacture our products in CPF (Cell Processing Facility) and no detection of endotoxin, mycoplasma and microorganisms in this area.)

N520 產品都是訂購後生產 交期約2-3週  效期約一年
一組kit 可以做2 test

Content of N520 is as below, for 2 patients
NK Cell Subculture Medium N201 1000 mL x2
NK Cell Initial Flask (M) N104 1 flask (75 cm2) x2
NK Cell Initial Medium N111a 90 mL x1
NK Cell Initial Cocktail N111b 3.8 mL x1

產品特色

・無需使用飼養細胞，即可從人外周血單核細胞（PBMC）擴增自然殺傷（NK）細胞。

・在2至3週的培養中，NK細胞可以擴增數百至數千倍。

・一個試劑盒足以使用20-50 mL全血擴增NK細胞患者。

・僅供研究使用。

(MDPI國際期刊) Phase I Clinical Trial Using Autologous Ex Vivo Expanded NK Cells and Cytotoxic T Lymphocytes for Cancer Treatment in Vietnam
https://www.mdpi.com/1422-0067/20/13/3166/htm
file:///C:/Users/Jimmy/Downloads/ijms-20-03166.pdf (下載點)

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)

• 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.

References

鄧学文、芦葉恵介、照沼篤、高根翼、鮫島葉月、贄田美江、照沼裕: 培養NK細胞の投与による体内NK細胞の活性化. *Medical

• Science Digest* 41, 42-45, 2015.
• 鄧学文、芦葉恵介、照沼篤、高根翼、鮫島葉月、贄田美江、照沼裕: 培養NK細胞の投与による体内NK細胞の活性化. *Medical
  Science Digest* 41, 42-45, 2015.
• Deng X, Terunuma H, Terunuma A, Takane T, Nieda M: Ex vivo-expanded natural killer cells kill cancer cells more effectively than ex vivo-expanded gd T cells or ab T cells. Int Immunopharmacol 22: 486-491, 2014
• Terunuma H, Deng X, Nishino N, Watanabe K: NK cell-based autologous immune enhancement therapy (AIET) for cancer. Stem Cells Regenerative Med 9: 9-13, 2013
• Deng X, Terunuma H, Nieda M, Xiao W, Nicol A: Synergistic cytotoxicity of ex vivo expanded naturel killer cells in combination with monoclonal antibody drugs against cancer cells. Int Immunopharmacol 14: 593-605, 2012