A kind of immune cell and preparation method thereof
An immune cell and cell technology, applied in the biological field, can solve the problems of low content of tumor antigens, insignificant effects, and low tumor antigen characteristics, and achieve strong tumor antigen sensitization, high affinity, and strong antigen sensitization. effect of action
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Embodiment 1
[0079] Example 1 Preparation of immune cells
[0080] (1) Culture of tumor cells and preparation of preparation 1
[0081] a, Tumor tissue separation Tumor cell A431 (adhesive growth tumor cell), 1640 medium+5% serum culture, inoculation density 5-10×10 5 cell / mL was inoculated in a T75 culture flask.
[0082] b. Change the liquid at half the amount the next day. The specific operation steps are as follows:
[0083] After the adherent growth tumor cells were 80% confluent, they were digested with 0.25% trypsin for 1-2 minutes, the serum was terminated, centrifuged at 400g for 5 minutes to pellet the cells, and subcultured at 1:2. Change the medium according to the cell growth status (change the medium once every 2-3 days), and collect the supernatant. After culturing for 4 days, all supernatants were collected, digested with 0.25% trypsin for 1-2 min, serum was terminated, centrifuged at 400 g for 5 min to collect cells.
[0084] c. The supernatant is concentrated by an ul...
Embodiment 2
[0102] Example 2 Preparation of immune cells
[0103] (1) Culture of tumor cells and preparation of preparation 1
[0104] a, Tumor tissue separation Tumor cells HL60 (suspension growth tumor cells), 1640 medium + 5% serum culture, inoculation density 5-10 × 10 5 cell / mL was inoculated in a T75 culture flask.
[0105] b. Change the liquid at half the amount the next day. The specific operation steps are as follows:
[0106] Rehydrate once every 2-3 days to maintain the cell density at 1-1.5×10 6 cell / mL, after 4-8 days, collect the cell suspension, centrifuge at 200-400g for 5min, and collect the supernatant and cells respectively.
[0107] c. The supernatant is concentrated by an ultrafiltration system, and the instrument uses a Sartorius Slice200 tangential flow device. Steps:
[0108] 1) The culture solution is pre-filtered through a filter membrane with a pore size of 0.45 μm to remove bulky impurities;
[0109] 2) Filter again through a filter membrane with a pore si...
Embodiment 3
[0124] Example 3 Preparation of immune cells
[0125] (1) Culture of tumor cells and preparation of preparation 1
[0126] a, Tumor tissue separation Tumor cell K562 (adhesive growth tumor cell), 1640 medium+5% serum culture, inoculation density 5-10×10 5 cell / mL was inoculated in a T75 culture flask.
[0127] b. Change the liquid at half the amount the next day. The specific operation steps are as follows:
[0128] After 90% of the adherent tumor-like cells were confluent, they were digested with 0.25% trypsin for 1-2 minutes, and the digestion was terminated with serum, centrifuged at 400g for 5 minutes to pellet the cells, and then subcultured at 1:2. Change the medium according to the cell growth status (change the medium once every 2-3 days), and collect the supernatant. After culturing for 8 days, all the supernatants were collected, digested with 0.25% trypsin for 1-2 min, the digestion was terminated by serum, and the cells were collected by centrifugation at 200 g ...
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