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Method for separating immune cells from blood and application of method to disease treatment

A technology of immune cells and cells, applied in the field of immune cell therapy for patients, can solve the problems of difficult blood immune cells, different methods of separation and expansion of blood immune cells, and complicated steps.

Active Publication Date: 2014-04-09
深圳博雅感知药业有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] However, at present, there are different methods for the separation and expansion of blood immune cells, and most of the steps are complicated. It is also difficult to obtain a large number of blood immune cells. At the same time, there are certain risks in collecting blood when patients suffer from diseases.

Method used

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  • Method for separating immune cells from blood and application of method to disease treatment
  • Method for separating immune cells from blood and application of method to disease treatment
  • Method for separating immune cells from blood and application of method to disease treatment

Examples

Experimental program
Comparison scheme
Effect test

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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Abstract

The invention relates to a simple and effective method for extracting blood immune cells, a cryopreservation and resuscitation method used with the former method and a method for proliferating the immune cells after resuscitation. The methods include the steps of whole-blood sample pretreatment, Ficoll separation, MNC (mononuclear cell) collection and washing, MNC counting, MNC cryopreservation, cell resuscitation and cell proliferation. By the aid of extraction, cryopreservation, resuscitation and proliferation with specific operation steps, the immune cells can be effectively obtained and used for treating tumors.

Description

technical field [0001] The present invention relates to a method for isolating immune cells from blood and its application in treating diseases, in particular to isolating and freezing immune cells from the blood of a healthy human body, and conducting immune cells to A method for in vitro expansion and administration of immune cells to patients. 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 self-replication characteristics. Lymphocytes have the characteristics of natural killing and self-protection, and play a key role in the treatment of viral, bacterial, and...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/078C12N5/0784C12N5/0783A01N1/02A61K35/14A61P35/00
Inventor 姚静杨传庆张辉郑爱玲张星星朱晓青
Owner 深圳博雅感知药业有限公司
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