Method for preparing cell group containing cytokine-induced killing cell
A technology of killing cells and cytokines, applied in animal cells, vertebrate cells, cell culture active agents, etc., can solve the problems of complicated collection operation and small number of cells, and achieve the effect of high-efficiency treatment effect.
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Embodiment 1
[0069] Example 1 Scaled-up culture of CIK cells using fibronectin fragment CH-296
[0070] (1) Isolation of PBMC
[0071] After the blood components were collected from 5 patients who gave informed consent (2 for kidney cancer, 1 for lung cancer, 1 for liver cancer, and 1 for thyroid cancer), the cell fraction containing PBMC was superimposed and centrifuged on Ficoll, and the peripheral blood mononuclear cells in the middle layer were recovered (PBMC).
[0072] (2) Immobilization of anti-human CD3 antibody and CH-296
[0073] Into a T225 flask was added a phosphate-buffered physiological solution containing an anti-human CD3 antibody at a final concentration of 5 μg / mL and CH-296 (Lacronac (registered trademark), manufactured by Takara Biotechnology Co., Ltd.) at a final concentration of 15 μg / mL. 40 mL of saline (PBS) or 40 mL of PBS containing only anti-human CD3 antibody at a final concentration of 5 μg / mL, and then incubated overnight at 4°C. PBS containing the antibod...
Embodiment 2
[0087] Example 2 Anti-tumor effect of CIK cells
[0088] (1) Transplantation of CIK cells into cancer patients
[0089] To each cancer patient, the self-derived R-CIKs prepared in Example 1 were implanted twice in 4 days as a round of administration, with an average administration of 3.9×10 10 indivual. All patients received at least 2 rounds of dosing, and one patient received 3 rounds of dosing.
[0090] (2) Anti-tumor effect
[0091] No serious adverse signs associated with cell administration were found in all R-CIKs-administered cancer patients.
[0092] By administration of R-CIKs, improvement of clinical symptoms and recovery of physiological functions were found in all patients. In addition, a significant decrease in calcitonin, a tumor marker, was found in patients with thyroid cancer. In addition, the tumor size of lung cancer patients was measured by PET-CT, and it was found that the tumor shrunk by about 50%.
[0093] In summary, R-CIKs is a very safe cell pr...
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