Method for separating mononuclear cells from whole blood collection device for blood donation

A collection device and nuclear cell technology, applied in the field of cell therapy products, to achieve the effects of promoting development, high enrichment, and sufficient sources

Pending Publication Date: 2022-05-31
QINGDAO BLOOD CENT
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Although this method uses passive immunotherapy, it is specific for killing tumor cells. Monoclonal antib...

Method used

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  • Method for separating mononuclear cells from whole blood collection device for blood donation
  • Method for separating mononuclear cells from whole blood collection device for blood donation
  • Method for separating mononuclear cells from whole blood collection device for blood donation

Examples

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Embodiment 1

[0040] A method for isolating peripheral blood mononuclear cells from a whole blood collection device for voluntary blood donation, comprising the following steps:

[0041] (1) Separation white filter:

[0042] Put the whole blood collection device for voluntary blood donation in a biological safety cabinet, spray 75% alcohol on the surface of the white filter and the tube walls on both sides for surface disinfection; clamp the tube walls on both sides with tube clamp 6, and cut it with sterile scissors 7 The upper and lower ends of the pipe walls on both sides have a long pipe reserved for the pipe walls on both sides, with a length of at least 15 cm.

[0043] (2) Forward and reverse flushing extraction white filter:

[0044] After cutting the side pipe connected with the collection bag, the reserved long pipe is located at the top, such as diagram 2-1 As shown, hang the white filter in the direction of the arrow downward, and use a 50mL syringe to wash the white filter pl...

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Abstract

The invention discloses a method for separating peripheral blood mononuclear cells from a repayment-free blood donation whole blood collection device, which comprises the following steps: clamping two side tube walls by using a tube clamp under a sterile condition, cutting open upper and lower end tube orifices of the two side tube walls by using sterile scissors, and reserving a section of long tube on each of the two side tube walls; using an injector to forward flush the white filter and backward flush the white filter with the flushing fluid, collecting the liquid obtained by backward flushing, centrifuging, sucking and discarding the supernatant after centrifuging, re-suspending cells with normal saline, adding into another two centrifugal tubes filled with the same amount of Ficoll, and continuing centrifuging; and after centrifugation is finished, sucking and abandoning supernatant, sucking the albuginea layer above the Ficoll into a new centrifuge tube, supplementing normal saline, sucking and abandoning supernatant after centrifugation, and manually counting by adopting a blood cell counting plate after the normal saline is resuspended. According to the method disclosed by the invention, the mononuclear cells can be aseptically extracted from the white filter of a blood donation and collection pipeline and cultured in vitro, and the cells cultured in vitro are used as DC-CIK treatment cells.

Description

technical field [0001] The invention relates to a method for separating peripheral blood mononuclear cells from a whole blood collection device for voluntary blood donation, in particular to a laboratory aseptic operation technique for obtaining human peripheral blood mononuclear cells on a large scale, which is used for subsequent The cell therapy and other uses belong to the technical field of cell therapy products. Background technique [0002] In the past ten years, cell therapy has developed rapidly as the frontier of medical development. The most important development direction of cell therapy, DC-CIK (dendritic cell-cytokine-induced killer cells) treats the regression of internal cancerous cells in the body, and uses activated immune Cells to eliminate cancerous autologous cells, through the co-culture of peripheral blood-derived dendritic cells (DC) and CIK cells, the most powerful antigen-presenting cells in the body, to obtain CLK cells stimulated by DC cells, and ...

Claims

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Application Information

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IPC IPC(8): C12N5/078
CPCC12N5/0634C12N2509/10
Inventor 李继明李蓓冯智慧安润
Owner QINGDAO BLOOD CENT
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