Resuscitation method of CAR-T cells
A technology of cells and resuscitation fluid, applied in the field of CAR-T cells, to achieve good survival rate, facilitate industrial use, and optimize the composition of resuscitation fluid
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Embodiment 1
[0023] A recovery method for CAR-T cells, comprising the following steps:
[0024] Take out the cryopreservation bag containing CAR-T cells and cryopreservation solution from liquid nitrogen, add resuscitation solution preheated at 36-40°C, shake the cryopreservation bag in a water bath at 36-40°C for 1-2 minutes, and centrifuge at 500rpm for 1 ~2min, discard the supernatant; add resuscitation solution preheated at 36~40℃, centrifuge at 500rpm for 1~2min, discard the supernatant, and resuspend the cells with RPMI-1640 medium; 2 to 3 times the volume of the freezing solution.
[0025] The resuscitation solution includes the following components: L-proline 50 μg / L, sodium selenite 10 mg / L, sodium pyruvate 2.2 mg / L, NaOH 0.2 mg / L, magnesium chloride 0.2 mg / L, astragaloside 4 25 mg / L L, Atractylodes lactone II 10mg / L, sodium diatrizoate 30mg / L, and the balance is RPMI-1640 medium.
Embodiment 2
[0027] A recovery method for CAR-T cells, comprising the following steps:
[0028] Take out the cryopreservation bag containing CAR-T cells and cryopreservation solution from liquid nitrogen, add resuscitation solution preheated at 36-40°C, shake the cryopreservation bag in a water bath at 36-40°C for 1-2 minutes, and centrifuge at 600rpm for 1 ~2min, discard the supernatant; add resuscitation solution preheated at 36~40℃, centrifuge at 600rpm for 1~2min, discard the supernatant, and resuspend the cells with RPMI-1640 medium; 2 to 3 times the volume of the freezing solution.
[0029] The resuscitation solution includes the following components: L-proline 50 μg / L, sodium selenite 10 mg / L, sodium pyruvate 2.2 mg / L, NaOH 0.2 mg / L, magnesium chloride 0.2 mg / L, astragaloside 4 25 mg / L L, Atractylodes lactone II 10mg / L, sodium diatrizoate 30mg / L, and the balance is RPMI-1640 medium.
Embodiment 3
[0031] The difference between this example and embodiment 1 is:
[0032] The resuscitation fluid includes the following components: L-proline 25 μg / L, sodium selenite 50 mg / L, sodium pyruvate 2.2 mg / L, NaOH 0.5 mg / L, magnesium chloride 0.4 mg / L, astragaloside IV 20 mg / L L, Atractylodes Ⅱ 25mg / L, sodium diatrizoate 18mg / L, and the balance is RPMI-1640 medium.
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