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Method for rapidly and efficiently amplifying tumor infiltrating lymphocytes

A technology of tumor infiltration and lymphocytes, applied in animal cells, vertebrate cells, blood/immune system cells, etc., can solve the problems of poor prognosis and ineffective treatment effect of liver cancer patients

Inactive Publication Date: 2013-11-20
爱瑞康医疗投资管理(北京)有限公司
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  • Description
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  • Application Information

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Problems solved by technology

Another study found that the effect of TIL adoptive cell therapy on other tumors such as liver cancer is not obvious. In liver cancer, tumor infiltrating CD8+ T cells express PD-1, which leads to poor prognosis of liver cancer patients, and PD-1 expression is correlated with Foxp3+ Treg infiltration. Related, high density of Foxp3+ Treg cells is an indicator of poor prognosis after hepatectomy

Method used

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  • Method for rapidly and efficiently amplifying tumor infiltrating lymphocytes
  • Method for rapidly and efficiently amplifying tumor infiltrating lymphocytes
  • Method for rapidly and efficiently amplifying tumor infiltrating lymphocytes

Examples

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Embodiment 1

[0038] The patient, female, 33 years old, the liver tumor tissue was fully washed with normal saline, and the normal tissue and necrotic area around the liver cancer specimen were removed. If overnight reprocessing is required, the liver cancer specimen should be transferred to a 50ml centrifuge tube containing about 10ml of complete medium and stored at 4°C. Put the liver cancer specimen in a sterile glass culture dish (Petri dish), add a small amount of normal saline, and use surgical scissors to cut out a size of about 1-2mm from different areas around the specimen 3 The number of small tissue blocks is generally not less than 8. Take a small piece of liver cancer tissue cut out and place it in one well of a 24-well plate, each well containing 2ml of complete medium and 6000 (or 3000) IU / ml of IL-2, and operate the other wells in the same way. Place the 24-well plate at 37°C, 5% CO 2 cultured in a cell culture incubator. The proliferation of lymphocytes around the tissue...

Embodiment 2

[0040] Patient, male, 72 years old, TIL culture method is consistent with the above method.

Embodiment 3

[0042] The patient was a 71-year-old male, and the TIL culture method was consistent with the above method.

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Abstract

The invention discloses a method for rapidly and efficiently amplifying tumor infiltrating lymphocytes. The method is mainly used for efficiently and rapidly amplifying tumor infiltrating lymphocytes (TIL) within a short period of time and basically realizing no influences on the phenotypes of the TIL. The method comprises: carrying out tumor tissue treatment, culturing liquid use, and IL-2 and OKT3 dosage and culturing treatment; taking 1-2mm<3> tissues from different areas of a liver cancer specimen, putting the tissues in the pores of a 24-pore plate, placing the pore plate in a culturing case, culturing, observing every other day, carrying out semis liquid replacement of the pore plate culturing liquid after the 5-6 day culture, carrying out semis liquid replacement of the pore plate culturing liquid every one or two days according to the TIL growth condition, collecting the TIL in all the pores once the 24-pore plate is full of the TIL, and culturing the TIL to form young TIL; and carrying out rapid in-vitro rapid amplification of the young TIL above 4000 times.

Description

technical field [0001] The invention relates to a method for rapidly and efficiently expanding tumor-infiltrating lymphocytes, more specifically, it relates to a method for culturing cells in adoptive immunotherapy, Background technique [0002] Tumor is one of the major diseases threatening human health. At present, malignant tumors are the second deadliest disease in the world and the first cause of death among urban and rural residents in China. The total number of new cancer patients in China is about 2 million every year, and the annual growth rate of new cancer patients is about 10%. Although surgery, radiotherapy and chemotherapy are still the main treatment methods for tumors, even radical surgery can only solve local problems and cannot avoid systemic metastasis. The biggest problem with radiotherapy and chemotherapy is that its killing effect is not targeted. Long-term chemotherapy and radiotherapy will damage the body's immune system and the functions of various ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/0783
Inventor 陈晚华孙庆文曲平波王延春颜廷骥李明张俊艺
Owner 爱瑞康医疗投资管理(北京)有限公司
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