Induced amplification method of CIK2 (NK NK-T) cells
A cell and immune cell technology, applied in the field of induction and expansion of CIK2 cells, can solve the problems of low expansion rate, difference in curative effect, and low content, and achieve the effect of high expression and strong efficacy
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Embodiment 1
[0061] (1) Induction and activation of immune cells (CIK2):
[0062] The induction and activation of immune cell CIK2 (NK NK-T) includes the following steps:
[0063] S01: Draw 50ml of anticoagulated peripheral blood Steps: Use 12,500 units of heparin sodium 1ml for anticoagulation, excluding sodium citrate for anticoagulation;
[0064] S02: Coat the T75 culture bottle with 7 ml of "CD16 monoclonal antibody (25-35 μg), Herceptin (trastuzumab 1-2 mg)" inducer for 24 hours, and suck off the coating solution for later use;
[0065] S03: 50ml of peripheral blood is separated to obtain 30ml of plasma (no need to inactivate);
[0066] S04: 3-5×10 lymphocytes isolated from peripheral blood 7 Put it into a coated T75 culture bottle, add 30ml of complete culture solution: AIM-V serum-free medium contains recombinant human γ-interferon (IFN-r) for injection, recombinant human interleukin-2 (IL-2) for injection , recombinant human interleukin-15 (IL-15), recombinant human interleukin-...
Embodiment 2
[0083] (1) Induction and activation of immune cells (CIK2):
[0084] The induction and activation of immune cell CIK2 (NK NK-T) includes the following steps:
[0085] S01: Draw 50ml of peripheral blood for anticoagulation: Use 12,500 units of heparin sodium 1ml for anticoagulation, excluding sodium citrate for anticoagulation;
[0086] S02: Coat the T75 culture bottle with 7 ml of "CD16 monoclonal antibody 25-35 μg, Herceptin (trastuzumab) 1-2 mg" inducer for 24 hours, suck off the coating solution and set aside;
[0087] S03: 50ml of peripheral blood separated and plasma (no inactivation required);
[0088] S04: Lymphocytes (3-5×10 7 Put into the coated T75 culture flask, add
[0089]Complete culture solution 30ml: AIM-V serum-free medium contains recombinant human γ-interferon (IFN-r) for injection, recombinant human interleukin-2 (IL-2) for injection, recombinant human interleukin-15 (IL-15 ), recombinant human interleukin-21 (IL-21), recombinant human stem cell growth ...
Embodiment 3
[0106] (1) Induction and activation of immune cells (CIK2):
[0107] The induction and activation of immune cell CIK2 (NK NK-T) includes the following steps:
[0108] S01: Take 50ml of peripheral blood for anticoagulation: use 12500 units of heparin sodium for anticoagulation, and exclude sodium citrate for anticoagulation;
[0109] S02: Coat the T75 culture bottle with 7 ml of "CD16 monoclonal antibody 25-35 μg, Herceptin (trastuzumab) 1-2 mg" inducer for 24 hours, suck off the coating solution and set aside;
[0110] S03: 50ml of peripheral blood separated and plasma (no inactivation required);
[0111] S04: Lymphocytes (3-5×10 7 Put it into a coated T75 culture flask, add 30ml of complete culture solution: AIM-V serum-free medium contains recombinant human γ-interferon IFN-r for injection, recombinant human interleukin-2 (IL-2) for injection, recombinant Human interleukin-15 (IL-15), recombinant human interleukin-21 (IL-21), recombinant human stem cell growth factor (SCF...
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