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Method for extracting PBMC in leucocyte film layer filter

An extraction method and buffy coat technology, applied in the field of biomedicine, can solve the problem of ineffective utilization of white blood cells and other problems, and achieve the effect of preventing cells from being destroyed

Inactive Publication Date: 2019-11-26
四川厚浦生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] There is no effective way and way to use the discarded leukocytes, and only a few literatures propose to obtain a small amount of leukocytes by washing the filter with PBS buffer

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] A kind of extraction method of PBMC in the buffy coat filter, its extraction method comprises:

[0022] (1) Wipe the leukocyte-removing filter and the outer walls of the pipelines on both sides with 75% ethanol, and wipe it twice;

[0023] (2) Clamp the pipes on both sides of the filter with sterile hemostatic forceps, quickly cut off the hose with sterile surgical scissors, and install a 50ml sterile needle-free syringe on the blood separation outlet side of the filter;

[0024] (3) Inject 200mll of flushing fluid into the filter and collect the flushing fluid that flows out. During the flushing process, place the filter horizontally, slowly inject the flushing fluid along the side of the blood separation outlet, flush in reverse, and collect the flushing fluid at the entrance of the blood separation direction;

[0025] (4) Finally, inject 50ml of air, drain the remaining liquid in the filter, and combine the collected flushing liquid;

[0026] (5) Use human lymphocyt...

Embodiment 2

[0029] A kind of extraction method of PBMC in the buffy coat filter, its extraction method comprises:

[0030] (1) Wipe the leukocyte-removing filter and the outer walls of the pipelines on both sides with 75% ethanol, and wipe it 3 times repeatedly;

[0031] (2) Clamp the pipes on both sides of the filter with sterile hemostatic forceps, quickly cut off the hose with sterile surgical scissors, and install a 50ml sterile needle-free syringe on the blood separation outlet side of the filter;

[0032] (3) Inject 300ml of flushing fluid into the filter and collect the outflowing flushing fluid. During the flushing process, place the filter horizontally, slowly inject the flushing fluid along the side of the blood separation outlet, and flush in the reverse direction. Collect the flushing fluid at the entrance of the blood separation direction. The proportioning (measured by volume) of each component is: 85% normal saline, 3% insulin, 2% transferrin, 1.5% selenium, 2.5% human albu...

Embodiment 3

[0037] A kind of extraction method of PBMC in the buffy coat filter, its extraction method comprises:

[0038](1) Wipe the leukocyte-removing filter and the outer walls of the pipelines on both sides with 75% ethanol, and wipe it twice;

[0039] (2) Clamp the pipes on both sides of the filter with sterile hemostatic forceps, quickly cut off the hose with sterile surgical scissors, and install a 50ml sterile needle-free syringe on the blood separation outlet side of the filter;

[0040] (3) Inject 250ml of flushing fluid into the filter and collect the flushing fluid that flows out. During the flushing process, place the filter horizontally, inject the flushing fluid slowly along the side of the blood separation outlet, and flush in reverse, and collect the flushing fluid at the entrance of the blood separation direction. The proportioning (measured by volume) of each component is: 85% normal saline, 3% insulin, 2% transferrin, 1.5% selenium, 2.5% human albumin, 1.5% interleuki...

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PUM

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Abstract

The invention discloses a method for extracting PBMC in a leucocyte film layer filter. The leucocyte film layer filter is washed repeatedly, leucocytes are separated and extracted from a washing fluid, discarded leucocytes in medical waste are recycled, a method for effectively recovering leucocytes from disposable leucocyte filters is established, the demands of base scientific research and evenclinical research can be met, and an important research material can be provided for immunology related research of disease treatment.

Description

technical field [0001] The invention relates to the field of biomedicine, in particular to a method for extracting PBMC in a buffy coat filter. Background technique [0002] White blood cells are one of the main components of blood. The main categories include: neutrophils, eosinophils, basophils, monocytes, and lymphocytes. Each type of white blood cell plays its own role in the immune system. However, if white blood cells enter the patient's body during blood transfusion, it may cause a transfusion reaction, so white blood cells are a waste product for clinical blood products. Clinical blood donations collected by medical institutions, as well as voluntary blood donations collected by blood stations, need to be separated from blood components before entering clinical reinfusion applications, and the white blood cells in the blood are removed through separation. At present, the commonly used blood component separation methods mainly include filtration method and separator...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/0786
CPCC12N5/0645C12N2509/00
Inventor 丁一
Owner 四川厚浦生物科技有限公司