Culture solution and culture method of DC cells
A technology of cell culture and culture medium, which is applied in the field of DC cell culture medium and its cultivation, can solve the problems of slow cell culture maturation time and low cell number purity, and achieve increased cell number and purity, good anti-tumor effect, and mature fast time effect
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Embodiment 1
[0041] Step 1, processing plasma to obtain leukocytes;
[0042] 1. Confirm that the suction tube of the plasma bag is sealed. Wipe the pipette and scissors with two different sterile room wipes soaked in 70% ethanol.
[0043] 2. Hold the suction tube with tweezers, cut the front end of the tube and control the flow rate with tweezers to transfer the plasma into a 50ml test tube. Record the plasma volume transferred.
[0044] 3. Centrifuge the tube (400g, 4°C) for 30 minutes to remove platelets.
[0045] 4. Pour the supernatant liquid into a new 50ml test tube.
[0046] 5. Put the test tube into a constant temperature water tank (56°C) and incubate for 30 minutes.
[0047] 6. Centrifuge tubes (400g, 4°C) for 60 minutes to remove precipitated fibrin.
[0048] 7. Pour the upper liquid (autologous serum) into a new 50ml test tube. The tubes were then labeled with the patient's name, date of preparation, and refrigerated (4°C).
[0049] 8. Confirm that the suction tube of the ...
Embodiment 2
[0090] Step 1, processing plasma to obtain leukocytes;
[0091] Same as embodiment 1;
[0092] Step 2, using adherent cell culture medium to cultivate leukocytes, and after culturing for 2 hours, remove the suspension, leaving adherent cells;
[0093] Adherent cell culture medium includes: 20ml of ALyS505N-0 culture medium, 9.9ul of GM-SCF with a concentration of 800U / ml, 6.3ul of IL-13 with a concentration of 500U / ml, and 50ul of 200mM L-glutamine.
[0094] Except that the adherent cell culture medium is different, others are the same as in Example 1;
[0095] Step 3, first wash the adherent cells with normal saline for 1-3 times, then add the subsequent first addition of DC cell culture medium to the culture flask with adherent cells, and culture for 20 hours at 36-38°C, 3-8% CO 2 ;
[0096] The subsequent first addition of DC cell culture medium includes: 15ml of serum-free DC medium, 10.3ul of GM-CSF at a concentration of 800U / ml, 6.7ul of IL-4 at a concentration of 500...
Embodiment 3
[0104] Step 1, processing plasma to obtain leukocytes;
[0105] Same as embodiment 1;
[0106] Step 2, using adherent cell culture medium to cultivate leukocytes, and after culturing for 2 hours, remove the suspension, leaving adherent cells;
[0107] Adherent cell culture medium includes: 20ml of ALyS505N-0 culture medium, 9.7ul of GM-SCF with a concentration of 800U / ml, and 6.6ul of IL-13 with a concentration of 500U / ml.
[0108] Except that the adherent cell culture medium is different, others are the same as in Example 1;
[0109] Step 3, first wash the adherent cells with normal saline for 1-3 times, then add the subsequent first addition of DC cell culture medium to the culture flask with adherent cells, and culture for 20 hours at 36-38°C, 3-8% CO 2 ;
[0110] The subsequent first addition of DC cell culture medium includes: 15ml of serum-free DC medium, 10.1ul of GM-CSF at a concentration of 800U / ml, 6.9ul of IL-4 at a concentration of 500U / ml, and 3.1ul of a concen...
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