Method for culturing cells B for long time
A B cell and cell technology, applied in the field of cell culture, can solve the problems of limited B cell proliferation multiple, cannot meet clinical use, and difficult to source, and achieve the effect of enhancing phagocytosis and killing effect, reducing risk, and multiplying multiple multiples.
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Embodiment 1
[0035] Medium formula:
[0036]
[0037]
[0038] (1) Collect 20ml of peripheral blood from healthy people;
[0039] (2) Collect plasma after centrifugation at 500g;
[0040] (3) Dilute the lower blood cells 1:1 with physiological saline, then add the diluted solution to the sepmate centrifuge tube (purchased from STEM CELL company) that has Ficoll separation solution (purchased from STEM CELL company), and centrifuge for 10-20 min;
[0041] (4) Pour the upper liquid from the centrifuge tube after centrifugation, and add RPMI 1640 medium to resuspend the cells. The amount of medium is determined by the number of cells, and the cell density is greater than 1×10. 7 / ml, centrifugal counting detection flow data;
[0042] (5) Take 1×10 among them 7 Cells, according to 1×10 6 The cell density of cells / ml is added to the culture medium of the present invention at 37°C, 5% CO2 for culture, and the fluid is supplemented every 3-4 days to ensure that the cell density is 1×10 6 Pcs / ml.
[0043] (6...
Embodiment 2
[0046] Medium formula:
[0047]
[0048]
[0049] (1) Collect 20ml of peripheral blood from healthy people;
[0050] (2) Collect plasma after centrifugation at 500g;
[0051] (3) Dilute the lower layer blood cells 1:1 with physiological saline, and then add the diluent to the sepmate centrifuge tube (purchased from STEM CELL company) that has Ficoll separation solution (purchased from STEM CELL company), and centrifuge for 10-20 min;
[0052] (4) Pour the upper liquid from the centrifuge tube after centrifugation, and add RPMI 1640 medium to resuspend the cells. The amount of medium is determined by the number of cells, and the cell density is greater than 1×10. 7 / ml, centrifugal counting detection flow data;
[0053] (5) Take 1×10 among them 7 Cells, according to 1×10 6 Cell density per ml added to the culture medium of the present invention at 37℃, 5% CO 2 Incubate and replenish the fluid every 3-4 days to ensure that the cell density is 1×10 6 Pcs / ml.
[0054] (6) On the 21st day of ...
Embodiment 3
[0057] Medium formula:
[0058]
[0059]
[0060] (1) Collect 20ml of peripheral blood from healthy people;
[0061] (2) Collect plasma after centrifugation at 500g;
[0062] (3) Dilute the lower layer blood cells 1:1 with physiological saline, and then add the diluent to the sepmate centrifuge tube (purchased from STEM CELL company) that has Ficoll separation solution (purchased from STEM CELL company), and centrifuge for 10-20 min;
[0063] (4) Pour the upper liquid out of the centrifuge tube after centrifugation, and add RPMI 1640 medium to resuspend the cells. The amount of medium is determined by the number of cells, and the cell density is greater than 1×10. 7 / ml, centrifugal counting detection flow data;
[0064] (5) Take 1×10 among them 7 Cells, according to 1×10 6 The cell density of cells / ml is added to the culture medium of the present invention at 37°C, 5% CO2 for culture, and the fluid is supplemented every 3-4 days to ensure that the cell density is 1×10 6 Pcs / ml.
[0065]...
PUM
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