Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

Method for effectively culturing tumor infiltrating lymphocytes (TILs)

A lymphocyte and tumor infiltration technology, applied in anti-tumor drugs, animal cells, vertebrate cells, etc., can solve the problems of lack of specificity of TIL cells, slow cell proliferation, and limited clinical efficacy, so as to reduce the burden of hospitalization and reduce hospitalization. The effect of shortening time and enhancing clinical efficacy

Inactive Publication Date: 2011-09-07
宋鑫 +1
View PDF2 Cites 18 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006]The current traditional TIL cell culture method is limited by the size of the tumor, the degree of tumor infiltration and the amount of pleural and peritoneal fluid, so there are problems of slow cell proliferation and low proliferation rate; There are great differences in the number of cells cultured among different patients, and the cells of many patients cannot reach the number of cells required for clinical treatment in a short period of time after culture; the commonly used culture methods at home and abroad need 40-50 days, and the clinical application is greatly limited ; clinical efficacy is still limited due to the lack of specificity of cultured TIL cells

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Method for effectively culturing tumor infiltrating lymphocytes (TILs)
  • Method for effectively culturing tumor infiltrating lymphocytes (TILs)
  • Method for effectively culturing tumor infiltrating lymphocytes (TILs)

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] The patient, male, 70 years old, had right lung cancer and a large amount of pleural effusion. Transfer 800ml of pleural effusion to a centrifuge tube, centrifuge at 1500rpm × 8min, take the supernatant of pleural effusion, mix the sediment with serum-free medium, separate the cells with lymphocyte separation medium and centrifuge 800g×20min. Aspirate the buffy coat, centrifuge at 2000rpm×10min, add serum-free medium, centrifuge at 1600rpm×8min, discard the supernatant to obtain mononuclear cells. Adjust the number of cells to 1.5 x 10 6 / ml, inoculate with RetroNectin250 μl, CD 3 Antibody 100μl Coated 175cm 2 In a culture bottle, placed in a saturated humidity of 90%, a temperature of 37°C, and CO 2 5.0% cultured in the incubator. After culturing for 24 hours, add IL-2 1000U / ml, and continue to store at 90% saturated humidity, temperature 37°C, CO 2 5.0% incubator culture. On the third day, the TIL cells were subcultured with serum-free medium containing 1000 U...

Embodiment 2

[0032] The patient, male, 65 years old, had left lung cancer. The surgical tissue of the lung was taken and placed in a sterile tank, and the tumor specimen was washed 2 or 3 times with sterile saline containing gentamicin 100-160 IU / ml. Add 10-20ml of culture medium to a 10cm diameter petri dish, put a piece of 200 mesh metal mesh, cut the tumor tissue into small pieces and place on the metal mesh. Crush the tumor tissue block with a 20ML syringe core, discard the metal mesh, collect the serum-free medium in the culture dish (cells flow into the medium to form a single-cell suspension), add the cell suspension to 20ml serum-free medium, 300RPM×3min. Take the supernatant, 1500 RPM × 6min, the sedimentation is the single cell layer. Add serum-free medium and mix well, separate the cells with lymphocyte separation medium and centrifuge at 800g×20min. Aspirate the buffy coat, centrifuge at 2000rpm×10min, add serum-free medium, centrifuge at 1600rpm×8min, discard the supernatant...

Embodiment 3

[0034] The patient, female, 68 years old, had multiple bone metastases from gastric adenocarcinoma and a large amount of ascites. Aseptically draw 650ml of pleural fluid. Transfer 650ml of pleural effusion to a 50Ml centrifuge tube, centrifuge at 1800rpm × 6min, take the supernatant of pleural effusion, mix the sediment with serum-free medium, separate cells with lymphocyte separation medium, and centrifuge at 800g × 22min. Aspirate the buffy coat, centrifuge at 2000rpm×10min, add serum-free medium, centrifuge at 1500rpm×8min, discard the supernatant to obtain mononuclear cells. Adjust the number of cells to 2.0×10 6 / ml, inoculate with RetroNectin250μl, CD with serum-free medium containing 0.01ml / ml of pleural fluid supernatant 3 Antibody 100μl Coated 175cm 2 In a culture bottle, placed in a saturated humidity of 90%, a temperature of 37°C, and CO 2 5.0% cultured in the incubator. After culturing for 24 hours, add IL-2 1500U / ml to the cells, and continue to store at 90%...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

No PUM Login to View More

Abstract

The invention discloses a method for effectively culturing tumor infiltrating lymphocytes (TILs). The method comprises the following steps: extracting mononuclear cells from the pectoral ascite or tumor tissue of a patient, washing, inoculating into a culture bottle coated with recombinant human fibrin and cluster-of-differentiation-3 antibody, culturing for 24 hours, adding interleukin II, then using a serum-free culture medium containing the interleukin II and the supernatant pectoral ascite to enlarge and subculture every two or three days, and adding apoptosis-regulated protein survivin and mucoprotein-1 on the twelveth or thirteenth day to activate the killing activity; and harvesting cells on the fourteenth day. By adopting the method, the culture time, which is only 14 days, is greatly shortened, so that the hospital stays of the patient can be greatly shortened and the hospital burden of the patient can be reduced. Meanwhile, the problems that the cell proliferation speed is low and the proliferation ratio is low can be solved, thus the number of the cultured cells can meet the clinical requirement. The cultured TILs has strong cell specificity so as to enhance the clinical curative effect.

Description

technical field [0001] The invention belongs to the technical field of autologous immune cell therapy, and in particular relates to a method capable of effectively cultivating tumor infiltrating lymphocytes. Background technique [0002] Tumor is one of the major diseases that seriously endanger people's life and health. At present, malignant tumors are the second leading cause of death 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 average annual growth rate of new cancer patients is about 10%. Surgery, chemotherapy, and radiotherapy are still the main treatment methods for tumors. However, even radical surgery can only solve local problems and cannot avoid systemic metastasis. The biggest problem with chemotherapy and radiotherapy is that their killing effects are not targeted. Long-term chemotherapy and radiotherapy will damage the body's immune sy...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
IPC IPC(8): C12N5/078A61P35/00
Inventor 宋鑫杨金艳
Owner 宋鑫
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com