Method for separating immune cells from blood and application of method to disease treatment
A technology of immune cells and cells, which is applied in the field of immune cell therapy for patients, and can solve problems such as different separation and expansion methods of blood immune cells, complicated steps, and difficulties in blood immune cells
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Embodiment 1
[0115] Embodiment 1, the method for immune cell extraction and cryopreservation
[0116] The immune cell extraction method comprises the following steps:
[0117] (1) Whole blood sample pretreatment: Turn the blood bag upside down 4-5 times, mix the blood sample thoroughly, transfer the blood to a 50ml centrifuge tube, and calculate the blood transferred to the centrifuge container according to the blood volume of 150ml, each centrifuge container Contain 25ml of blood, 6 tubes in total, take 5ml of blood sample in a 15ml centrifuge tube for blood type testing, add 25ml of normal saline to a 50ml centrifuge tube for equal volume 1:1 dilution, mix gently and fully with a 25ml pipette uniform;
[0118] (2) Ficoll method separation: Calculated according to 150ml blood volume, take 12 50ml centrifuge tubes, add 15ml Ficoll cell separation solution to each tube, draw 25ml diluted blood sample with a 25ml pipette and slowly add it to Ficoll along the inclined tube wall For the up...
Embodiment 2
[0125] Embodiment 2, the method for immune cell extraction and cryopreservation
[0126] Carried out with reference to the method of Example 1. Volunteer Li Si (pseudonym), 41 years old, female, with a blood volume of 115ml. The total amount of mononuclear cells collected is calculated in step (4) 1.40×10 8 cells, converted into 100ml blood volume is calculated as 1.22×10 8 cells. Step (5) The number of frozen tubes is 12 tubes, of which at least 7 tubes are in accordance with 1×10 7 / ml density frozen.
Embodiment 3
[0127] Embodiment 3, the method for immune cell extraction and cryopreservation
[0128] Carried out with reference to the method of Example 1. Volunteer Wang Wu (pseudonym), 33 years old, male, blood collection volume 120ml. The total amount of mononuclear cells collected in step (4) is calculated to be 1.69×10 8 cells, converted into 100ml blood volume is calculated as 1.41×10 8 cells. Step (5) The number of frozen tubes is 14 tubes, of which at least 2 tubes are 5×10 6 / ml density frozen storage, at least 3 tubes according to 1×10 7 / ml density, at least 2 tubes of which are in accordance with 2×10 7 / ml density frozen.
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