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Process of caltivating TIL cell by self hydrothorax supernatant

A cell and pleural effusion technology, applied in tissue culture, biochemical equipment and methods, microorganisms, etc., can solve the problems of increasing the financial burden of patients, prolonging the treatment time of patients, increasing the probability of pollution, etc., to reduce the economic burden, reduce pain, and achieve effective results. Good results

Inactive Publication Date: 2005-01-05
THE AFFILIATED DRUM TOWER HOSPITAL MEDICAL SCHOOL OF NANJING UNIV
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AI Technical Summary

Problems solved by technology

One is that the in vitro culture time is too long, which increases the probability of contamination and cannot be reinfused; the other is that human AB serum is used in the serum medium, which has the potential to cause infectious diseases such as hepatitis B and AIDS; Human AB serum leads to increased cost, increases the financial burden of the patient, and also prolongs the treatment time of the patient

Method used

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Examples

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Embodiment Construction

[0013] Aseptically collect about 1000ml of cancerous pleural effusion from each patient, and immediately separate TIL from tumor cells in a 100-level laboratory. The membranous cells were cultured with the autologous supernatant reserved as the culture medium for 12 to 24 hours, usually 16 hours. It can be seen that all the tumor cells adhered to the wall, and the suspension cells were almost all TILs; the obtained TILs were divided into 1×10 6 / ml concentration suspended in the autologous supernatant, placed in 37 ° C, 5% CO 2 In the incubator; within 24 hours, that is, after one day, add OKT3 with a final concentration of 100ng / ml, IL-2 at 6000U / ml, and PHA at 100ug / ml; every 1 to 2 days, add IL- 2 complete medium. Then, the TIL obtained from the culture was reinfused into the patient's chest within 3 days to treat malignant pleural effusion.

[0014] The culture of TILs using autologous pleural fluid supernatant was compared with the culture of TILs in RPMI 1640 containin...

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PUM

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Abstract

The invention is a method of culturing TIL (tumour infiltration lymphocytes) by using self-pleural effusion supernatant. It centrifuges the collected malignant pleural effusion to store the supernatant as culture medium, places albugineous-layer cells in bottom-layer cells in the culture medium for culturing, so as to obtain suspended TIL, cultures the suspended TIL in a hatching box for 1 days, then adds in OKT3, IL-2, PHA, etc, and after 1 or 2 days, adds in IL-2 for culturing. The invention avoids the defects of existing methods: the infection of hepatitis B, AIDS, etc caused by using human AB serum, the pollution probability increased by overlong culturing time in vitro, increasing the burden of patients, prolonging curing time and increasing cost, etc.

Description

technical field [0001] The invention relates to a method for cultivating TIL cells, in particular to a method for cultivating TIL cells using autologous pleural effusion supernatant. Background technique [0002] Tumor infiltrating lymphocytes (tumor infiltrating lymphocytes, TIL for short) are the second generation of anti-tumor effector cells after LAK cells, which can specifically kill tumor cells, and their anti-tumor activity is 50-100 times stronger than that of LAK cells. One of the options for anti-tumor cell immunotherapy. [0003] Before the present invention was made, the method for preparing TIL was mainly to obtain TIL by discontinuous density gradient centrifugation, and then culture it in vitro with artificially synthesized medium, usually RPMI1640 medium containing 10% human AB serum, for about 2 weeks. Time, and then re-infused into the patient's chest to kill the tumor cells in the patient's body. This method of culturing TILs has many drawbacks. One is ...

Claims

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Application Information

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IPC IPC(8): C12N5/08
Inventor 刘宝瑞邹征云
Owner THE AFFILIATED DRUM TOWER HOSPITAL MEDICAL SCHOOL OF NANJING UNIV
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