Method for automatically separating immune cells and extracting PRP from adult peripheral blood
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A technology for automatic separation of immune cells, applied in the direction of blood/immune system cells, animal cells, vertebrate cells, etc., can solve the problem of no preparation method
Active Publication Date: 2015-06-17
FIVE DIMENSION BY INCOSC HEALTH MANAGEMENT JIANGSU
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[0008] So far, there is no uniform preparation method for PRP. The principle of PRP preparation is mainly to use the different sedimentation speeds of various components in the blood, and separate the blood into layers by centrifugation.
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Embodiment 1
[0116] Embodiment 1, the method for immune cell extraction and cryopreservation
[0117] The immune cell extraction method comprises the following steps:
[0118] (1) Whole blood sample pretreatment: Centrifuge the collected adult peripheral blood sample at 1000rpm for 10min; absorb the supernatant plasma to extract platelet-rich plasma; transfer the remaining lower layer to an AXP processing bag, add hydroxyethyl starch solution and place it Mix well on a shaker; the mixing volume ratio of blood sample to hydroxyethyl starch solution is 4:1; the concentration of hydroxyethyl starch solution is 6%, which is to add hydroxyethyl starch to 0.9% sodium chloride solution prepared in , and 0.75% maltose is added to the hydroxyethyl starch;
[0119] (2) Automatic separation: After putting the AXP processing bag into the AXP automatic cell separation equipment, automatic separation is performed; the automatic separation performs two centrifugation treatments, the first centrifugati...
Embodiment 2
[0134] Embodiment 2, the method for immune cell extraction and cryopreservation
[0135] 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 freeze.
Embodiment 3
[0136] Embodiment 3, the method for immune cell extraction and cryopreservation
[0137] 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 freeze.
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Abstract
The invention relates to a simple and effective method for automatically separating immune cells and extracting PRP from adult peripheral blood, in particular to a method for automatically separating immune cells and extracting platelet rich plasma (PRP) from the peripheral blood y using an AXP full-automatic cell separation device, a cryopreservation and resuscitation method used together with the method, and a method for amplifying the post-resuscitation immune cells. The method for automatically separating the immune cells and extracting PRP from the adult peripheral blood comprises the following steps: preprocessing whole-blood samples, automatically separating, collecting mononuclear cells, and optionally detecting one and more of the following projects on the obtained immune cells: the total number of the mononuclear cells, the living rate of the mononuclear cells, cell pollution, genetic diseases and HLA-ABC / DR matching. By virtue of the adoption of the special operation steps of extraction, cryopreservation, resuscitation and amplification, the immune cells can be effectively obtained and can be used for treating tumor; meanwhile, by virtue of the method, the platelet rich plasma can be effectively obtained, so that the effective clinical treatment methods can be provided.
Description
technical field [0001] The invention relates to a method for automatically separating immune cells from blood, especially peripheral blood, and simultaneously extracting platelet-rich plasma (PRP) and its application in treating diseases, in particular to automatically separating immune cells from healthy human blood, especially peripheral blood. Cells are frozen and stored, and when immunocompromised or disease symptoms occur, immune cells are expanded in vitro and administered to patients for immune cell therapy. Background technique [0002] Immune cell is a common name for white blood cells, including lymphocytes and various phagocytes, and specifically refers to lymphocytes that can recognize antigens and generate specific immune responses. Lymphocytes are the basic components of the immune system and are widely distributed in the body. Studies have found that blood is rich in lymphocytes, which have attracted people's attention because of their immune regulation and s...
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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/078C12N5/0786
Inventor 许晓椿李诣书周丹肖海蓉
Owner FIVE DIMENSION BY INCOSC HEALTH MANAGEMENT JIANGSU
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