Cell cryopreservation liquid and cell cryopreservation method
A cryopreservation method and cryopreservation solution technology, which is applied in the field of cell cryopreservation solution and cell cryopreservation, can solve the problems of high pollution risk and high price, and achieve the effects of low pollution risk, reduced damage, and improved survival rate
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Embodiment 1
[0064] 1. After culturing the 293 cells to the logarithmic phase, in a sterile environment, use an electric pipette to suck up the cell culture medium in the culture dish, add 10mL PBS buffer solution to the culture dish, shake gently to wash, The PBS buffer was then removed with an electric pipette;
[0065] 2. In a sterile environment, evenly add 0.5 mL of 0.25% trypsin solution to the surface of the 293 cells in the culture dish, digest in a 37°C incubator for 2 minutes, and add 3 mL of whole cell culture solution to the culture dish , to terminate the digestion of trypsin, and then use an electric pipette to blow off the 293 cells from the culture dish to suspend the 293 cells in the above-mentioned whole cell culture solution; wherein, the above-mentioned whole cell culture solution includes a volume ratio of 9:1 The cell culture medium and fetal calf serum, the cell culture medium includes the following components:
[0066] Anhydrous calcium chloride 265.00mg / L; ferric ...
Embodiment 2
[0072] 1. After culturing the MCF-7 cells to the logarithmic phase, in a sterile environment, use an electric pipette to suck up the cell culture medium in the culture dish, add 15mL PBS buffer solution to the culture dish, and shake gently. Wash, then remove the PBS buffer with an electric pipette;
[0073] 2. In a sterile environment, evenly add 2 mL of 0.25% trypsin solution to the surface of the MCF-7 cells in the culture dish, digest in a 37°C incubator for 5 minutes, and add 5 mL of whole cell culture to the culture dish solution to stop the digestion of trypsin, and then use an electric pipette to blow off the MCF-7 cells from the culture dish to suspend the MCF-7 cells in the above-mentioned whole cell culture solution; wherein, the above-mentioned whole cell culture solution includes volume Cell culture medium and fetal bovine serum in a ratio of 9:1, the cell culture medium comprising the following components:
[0074] Calcium chloride 116.6mg / L; copper sulfate 0.00...
Embodiment 3
[0080] 1. After culturing HL60 cells to the logarithmic phase, centrifuge the cell culture solution containing the logarithmic phase cells at 800 rpm for 6 minutes, discard the supernatant to obtain the HL60 cell pellet, and then add 12mL PBS buffer to resuspend the cells , then centrifuged at 800 rpm for 6 min, discarded the supernatant to obtain the washed HL60 cell pellet;
[0081] 2. Suspend the washed HL60 cell pellet with cell freezing solution, then count the cells, and dilute with the above cell freezing solution to obtain a concentration of 2×10 6A mixture of HL60 cells in logarithmic phase per milliliter and cell cryopreservation solution; wherein, the above-mentioned cell cryopreservation solution includes 90% whole cell culture solution and 10% dimethyl sulfoxide by volume percentage, and the whole cell The culture medium includes cell culture medium and fetal calf serum with a volume ratio of 9:1, and the cell culture medium includes the following components:
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