A kind of ready-to-use erythrocyte cryoprotectant and its application method
A cryoprotectant and red blood cell technology, applied in the field of cells, can solve the problems of complex freezing process, short storage time, long recovery time, etc., and achieve the effects of eliminating washing steps, short time-consuming and simple components
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Embodiment 1
[0023] (1) Add trehalose, human serum albumin, and DMSO to the aqueous solution in sequence to prepare a red blood cell cryoprotectant, in which the content of DMSO is 6% (v / v), and the concentration of trehalose is 0.28mol / L; human serum albumin The protein content is 2% (v / v);
[0024] (2) Centrifuge fresh blood with added anticoagulant at 4°C and 2500 rpm for 10 minutes, remove the plasma to obtain concentrated red blood cells, mix 10 mL of concentrated red blood cells with 100 mL of red blood cell cryoprotectant, place at room temperature for 30 minutes, and place in gradient In the cooling box, the temperature was gradually lowered to -80°C in a deep-low temperature refrigerator, and the cooling rate was 1°C / min. After 7 days, it was taken out and placed in a water bath at 37°C with constant shaking to recover.
[0025] The recovery rate of erythrocytes determined by hemocytometer counting was 54.69%, and the cell viability of erythrocytes measured by trypan blue detecti...
Embodiment 2
[0027] (1) Add trehalose, human serum albumin, and DMSO to the aqueous solution in sequence to prepare a red blood cell cryoprotectant, in which the content of DMSO is 5% (v / v), and the concentration of trehalose is 0.3mol / L; human serum albumin The protein content is 2% (v / v);
[0028] (2) Centrifuge fresh blood with added anticoagulant at 4°C and 2500 rpm for 10 minutes, remove the plasma to obtain concentrated red blood cells, mix 10 mL of concentrated red blood cells with 100 mL of red blood cell cryoprotectant, place at room temperature for 30 minutes, and place in gradient In the cooling box, the temperature was gradually lowered to -80 °C in a deep-low temperature refrigerator, and the cooling rate was 2 °C / min. After 7 days, it was taken out and placed in a water bath at 37°C with constant shaking to recover.
[0029] The recovery rate of erythrocytes determined by hemocytometer counting was 45.28%, and the cell viability of erythrocytes measured by trypan blue detect...
Embodiment 3
[0031] (1) Add trehalose, human serum albumin, and DMSO to the aqueous solution in sequence to prepare a red blood cell cryoprotectant, in which the content of DMSO is 7% (v / v), and the concentration of trehalose is 0.15mol / L; human serum albumin The protein content is 2% (v / v);
[0032] (2) Centrifuge the fresh blood with added anticoagulant at 4°C and 2500 rpm for 10 minutes, remove the plasma to obtain concentrated red blood cells, mix 10 mL of concentrated red blood cells with 50 mL of red blood cell cryoprotectant, place at room temperature for 30 minutes, and place in gradient In the cooling box, the temperature was gradually lowered to -80°C in a deep-low temperature refrigerator at a cooling rate of 1.5°C / min. After 7 days, it was taken out and placed in a 37°C water bath for recovery with constant shaking.
[0033] The recovery rate of erythrocytes determined by hemocytometer counting was 30.69%, and the cell viability of erythrocytes measured by trypan blue detection...
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