In-vitro culture kit for CMV-CTLs (human cytomegalovirus-cytotoxic T lymphocytes)

A CMV-CTL, in vitro culture technology, applied in the field of in vitro culture kits for CMV-CTL cells

Inactive Publication Date: 2017-06-16
广州可伴咨询中心(普通合伙)
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The live CMV vaccine developed abroad can induce antibodies, but the problem of excluding the carcinogenic potential of the vaccine remains to be resolved

Method used

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  • In-vitro culture kit for CMV-CTLs (human cytomegalovirus-cytotoxic T lymphocytes)
  • In-vitro culture kit for CMV-CTLs (human cytomegalovirus-cytotoxic T lymphocytes)
  • In-vitro culture kit for CMV-CTLs (human cytomegalovirus-cytotoxic T lymphocytes)

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0059] The kit of the present invention is a standardized and standardized instrument prepared for the in vitro culture method of CTL cells. The contents of the kit are described in detail below:

[0060] The contents of the kit include: sterile centrifuge tubes for holding liquids, 10×15mL / pcs, 17×50mL / pcs, 6×250mL / pcs; sterile pipettes for sucking and releasing liquids, 14× 10ml / bottle; cell sorting fluid for sorting cells, 10×5mL / bottle; coated bottle “CTL5” for inducing and culturing CTL cells, 1×175cm 2 / piece; Coated bottle "CTL6" for inducing and culturing T cells, 1×175cm 2 / piece; serum-free medium used to provide nutrients needed for growth of cells, 2×1000mL / bottle; sterile cell culture bag used as a container for culturing cells, 1×2000mL / piece; used to stimulate CTL, T cells Cytokine 9 for division and proliferation, namely IL-2, 1×500,000 IU / cartridge; cytokine 10 for stimulating CTL and T cell division and proliferation, namely IL-2, 1×1 million IU / cartridge;...

Embodiment 2

[0070] combine Figure 1-8 The present invention is described in detail, human peripheral blood is used as a processed sample, and the contents of the kit of the present invention are used to sort and induce CMV-CTL cells. The specific operation steps are as follows:

[0071] 1. Blood processing

[0072] 1. Pour 50mL of anticoagulated blood into a 50mL centrifuge tube.

[0073] 2. Slowly pour the blood sample into the cell sorting solution HISTOPAQUE-1077, 6mL per tube.

[0074] 3. Centrifuge at 550xg, 30min, 4°C.

[0075] 4. The cells in the centrifuge tube after centrifugation are divided into four layers from top to bottom. (First layer: plasma; second layer: ring milky white lymphocyte layer; third layer: clear cell sorting liquid layer; fourth layer: red blood cell layer.) Collect the first layer and transfer it to a new 50mL centrifuge tube , inactivated at 56°C for 30 minutes, which is autologous plasma; collect the second layer and transfer it into two new 50mL ce...

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Abstract

The invention relates to an in-vitro culture kit, in particular to an in-vitro culture kit for CMV-CTLs (human cytomegalovirus-cytotoxic T lymphocytes). The in-vitro culture kit comprises a CMV antigenic peptide coating bottle for inducing CTL cells to be differentiated, a coating bottle for inducing T cells to be differentiated, cell factors stimulating CTL and T cells to be split and proliferated as well as cell factors enabling associated antigens carried by the T cells to stimulate the CMV-CTLs to proliferate and mature. Complicated reagents and consumables required for in-vitro culture of the CMV-CTLs are standardly combined, all operation is standard and modeled, the working efficiency is improved, and the cell quality and yield differences brought by worker operation are reduced.

Description

technical field [0001] The invention relates to an in vitro culture kit, in particular to an in vitro culture kit for CMV-CTL (human cytomegalovirus-CTL) cells. Background technique [0002] Human cytomegalovirus (human Cytomegalovirus, HCMV, referred to as CMV), also known as cellular inclusion virus, belongs to the family Herpesviridae, and has a high infection rate in the population, and the infection rate in adults in developed countries is 40% to 60%. It can be as high as 100% in my country, and the adult infection rate in China is more than 95%. Usually, it is a recessive infection, and when the immunity is low (such as after organ transplantation, AIDS, etc.), it will be reactivated and cause severe infection. At present, there are at least 55,000 to 60,000 cases of allogeneic hematopoietic stem cell transplantation every year, and the number of transplants is increasing every year. CMV infection is one of the serious complications of hematopoietic stem cell transplant...

Claims

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Application Information

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IPC IPC(8): C12N5/0783
CPCC12N5/0638C12N2501/2302C12N2501/2315C12N2501/2321C12N2501/24C12N2501/515C12N2501/59C12N2501/599C12N2501/998
Inventor 邢威李佳
Owner 广州可伴咨询中心(普通合伙)
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