In-vitro culture kit for EBV-CTL

A technology of EBV-CTL and in vitro culture, applied in clinical application research and biological fields, can solve the problems of treatment failure of nasopharyngeal carcinoma

Inactive Publication Date: 2017-05-31
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

However, local recurrence and distant metastasis are still the main reasons for the failure of nasopharyngeal carcinoma.

Method used

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  • In-vitro culture kit for EBV-CTL
  • In-vitro culture kit for EBV-CTL
  • In-vitro culture kit for EBV-CTL

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0061] 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:

[0062] 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 solution for sorting cells, 10×5mL / bottle; EBV antigen peptide-coated bottle “CTL1” for inducing the differentiation of cultured CTL cells, 1×175cm 2 / piece; Coated bottle "CTL2" for inducing the differentiation of cultured 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 1 for division and proliferation, namely IL-2, 1×500,000 IU / cartridge; cytokine 2 for stimulating CTL and T cell division...

Embodiment 2

[0072] combine Figure 1-10 The present invention is described in detail. Human peripheral blood was used as a processed sample, and the induced EBV-CTL cells were sorted using the contents of the kit of the present invention. The specific operation steps are as follows: 1. Blood processing

[0073] 1. Pour 50mL of anticoagulated blood into a 50mL centrifuge tube.

[0074] 2. Slowly inject the blood sample onto the cell sorting medium, 6mL per tube.

[0075] 3. Centrifuge at 550xg, 30min, 4°C.

[0076] 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 centrifuge tubes, add s...

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Abstract

The invention relates to the technical fields of clinical application study and biology, and particularly relates to an in-vitro culture kit for inducing mononuclear cells obtained from human peripheral blood into nasopharyngeal carcinoma EB virus-CTL (EBV-CTL). The kit comprises a serum-free cell culture medium, a cell sorting solution for cell sorting, a cell dispersing agent for preventing cell aggregation, an EBV antigen peptide coating bottle for inducing CTL differentiation, a coating bottle for inducing T cell differentiation, a cell factor causing a T cell to carry a related antigen to stimulate cell proliferation, and a cell factor for stimulating differentiation and proliferation of CTL and T cells. According to the invention, complex reagents and consumables needed in in-vitro culture of EBV-CTL are combined normative, and all operations are normalized and modeled. Thus, the working efficiency is improved, and the difference in cell quality and yield caused by operations of the personnel can be reduced.

Description

technical field [0001] The invention relates to the fields of clinical application research and biotechnology, in particular to an in vitro culture kit for inducing EBV-CTL (nasopharyngeal carcinoma EB virus-CTL) from human peripheral blood mononuclear cells. Background technique [0002] According to a rough estimate by the United Nations Health Organization (WHO), about 80% of nasopharyngeal carcinoma (nasopharyngeal carcinoma, NPC) patients in the world are in China, and it is common in southern China, especially in Guangdong, where the incidence rate is relatively high (20-50 / 100 000 ), nasopharyngeal carcinoma is a tumor that deserves attention and attention. Due to the particularity of the location of nasopharyngeal carcinoma, there are many important organs at risk around it, and radical surgery cannot be performed. Therefore, radiotherapy and chemotherapy have become the first choice for the treatment of nasopharyngeal carcinoma. With the continuous advancement of r...

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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