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Method of automatically extracting immune cells in APB (Adult Peripheral Blood)

A technology of immune cells and peripheral blood, which is applied in the field of in vitro expansion and application to patients, and can solve the problems of different separation and expansion methods of blood immune cells, complicated steps, and difficulties in blood immune cells.

Active Publication Date: 2015-06-10
JIANGSU BOYA REGENERATIVE MEDICAL TECH
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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 of automatically extracting immune cells in APB (Adult Peripheral Blood)
  • Method of automatically extracting immune cells in APB (Adult Peripheral Blood)
  • Method of automatically extracting immune cells in APB (Adult Peripheral Blood)

Examples

Experimental program
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Effect test

Embodiment 1

[0101] Example 1. Methods for immune cell extraction and cryopreservation

[0102] The immune cell extraction method includes the following steps:

[0103] (1) Whole blood sample pretreatment: Transfer the collected adult peripheral blood blood sample to an AXP processing bag, add hydroxyethyl starch solution and place it on a shaker to mix thoroughly; the mixing volume ratio of blood sample to hydroxyethyl starch solution It is 4:1; the concentration of hydroxyethyl starch solution is 6%, which is prepared by adding hydroxyethyl starch to 0.9% sodium chloride solution, and 0.75% maltose is also added to the hydroxyethyl starch;

[0104] (2) Automatic separation: After the AXP treatment bag is put into the AXP automatic cell separation equipment, automatic separation is performed; the automatic separation is performed twice by centrifugation. Centrifugation conditions are: 80rcf, 10min, 15℃;

[0105] (3) Mononuclear cell collection: The immune cells automatically separated...

Embodiment 2

[0119] Example 2. Methods of Immune Cell Extraction and Cryopreservation

[0120] Refer to the method of Example 1. Volunteer Li Si (pseudonym), 41 years old, female, with a blood volume of 115ml. Step (4) Calculate the total amount of mononuclear cells collected 1.40 × 10 8 100ml of blood volume is calculated as 1.22×10 cells 8 cells. Step (5) The number of cryopreserved tubes is 12 tubes, of which at least 7 tubes are in accordance with 1×10 7 Cryopreserved at a density of / ml.

Embodiment 3

[0121] Example 3. Methods for immune cell extraction and cryopreservation

[0122] Refer to the method of Example 1. Volunteer Wang Wu (pseudonym), 33 years old, male, with a blood volume of 120ml. In step (4), the total amount of mononuclear cells collected is 1.69×10 8 100ml of blood is calculated as 1.41×10 cells 8 cells. Step (5) The number of cryopreserved tubes is 14 tubes, of which at least 2 tubes are in accordance with 5×10 6 Cryopreserved at a density of / ml, at least 3 tubes are in accordance with 1 × 10 7 Cryopreserved at a density of 7 Cryopreserved at a density of / ml.

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Abstract

The invention relates to a simple and effective method of automatically extracting immune cells in APB (Adult Peripheral Blood), in particular to a method of using an AXP full-automatic cell separation equipment to extract the immune cells in the APV, a supporting freezing-thawing method, and an immune cell expansion method after the thawing. The method comprises the steps of whole blood sample preprocessing, automatically separating, mononuclear cell collection and at least one of the following items of detecting the immune cells: total mononuclear cells, mononuclear cell viability, cell contamination, genetic disease, HLA-ABC / DR matching. According to the method provided by the invention, the immune cells can be effectively obtained by using the specific operating steps, such as extraction, freezing, thawing and amplification, and the immune cells can be used for treating tumors.

Description

technical field [0001] The present invention relates to a method for automatically separating immune cells from blood, especially peripheral blood, and its application in the treatment of diseases, in particular to automatically separating immune cells from healthy human blood, especially peripheral blood, and cryopreserving them, and to A method of expanding immune cells in vitro and administering them to a patient for immune cell therapy when the disease is low or symptomatic. Background technique [0002] Immune cell is a common name for white blood cells, including lymphocytes and various phagocytic cells, and also specifically refers to lymphocytes that can recognize antigens and produce specific immune responses. Lymphocytes are the basic components of the immune system and are widely distributed in the body. Studies have found that the blood is rich in lymphocytes. Because of the characteristics of immune regulation and self-replication, these cells have always attra...

Claims

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Application Information

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IPC IPC(8): C12N5/0783C12N5/0784
Inventor 许晓椿周丹王芳肖海蓉
Owner JIANGSU BOYA REGENERATIVE MEDICAL TECH
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