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Immune isolated cell medicine, its preparing process and its application in killing tumours

A tumor and drug technology, applied in the field of novel immune isolation cell drugs, to achieve good therapeutic effect, poor therapeutic effect, and less toxic and side effects

Inactive Publication Date: 2006-02-08
崔和厚
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, so far, there have been no reports of using APA microcapsules to encapsulate two or more tumor-suppressing and / or tumor-killing factor-secreting cells and implanting them in tumors for the treatment of tumors.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0050] Embodiment 1: The establishment of rEST / 293 genetic engineering cell line

[0051] 1. Transfect the expression vector of human vascular endostatin (rEST) into human embryonic kidney 293 cells to obtain engineered cells that stably secrete human vascular endostatin (rEST / 293)

[0052] 2. The viability of rEST / 293 cells was detected by the pan blue staining method;

[0053] 3. Detect the expression of endostatin in the transfected cells from the mRNA level by RT-PCR amplification method;

[0054] 4. Detect the expression of endostatin in transfected cells from the protein level: use SDS-PAGD to detect the specificity of rEST; use Western Blot to confirm that the secreted protein expressed by rEST / 293 cells contains recombinant rEST and has antigenic activity;

[0055] 5. The amount of rEST secreted by the cells was detected by ELISA.

[0056] 6. Identification of in vitro biological activity of endostatin expressed by transfected cells: Inhibition experiments of vascula...

Embodiment 2

[0057] Embodiment 2: the establishment of TNF / 293 genetic engineering cell line

[0058] 1. Transfect the tumor necrosis factor alpha (TNFα) expression vector into human embryonic kidney 293 cells to obtain engineered cells (TNFα / 293) that stably secrete human vascular endostatin;

[0059] 2. The survival rate of TNFα / 293 cells was detected by the pan blue staining method;

[0060] 3. Detect the expression of TNFα in transfected cells from the mRNA level by RT-PCR amplification method;

[0061] 4. Use Western Blot method to detect the expression of TNFα in transfected cells at the protein level;

[0062] 5. The amount of TNFα secreted by cells was detected by ELISA method.

[0063] 6. The killing activity of TNFα on tumor cells was determined by MTT colorimetry.

[0064] It should be noted that the methods of Examples 1 and 2 belong to conventional techniques, and it does not have any limiting effect on the present invention.

Embodiment 3

[0065] Embodiment 3: Preparation of APA-TIF cell microcapsule drug

[0066] 1. Use the TIF cell (rEST / 293 cell, TNFα / 293 cell mixture) precipitate obtained by the method in Example 1 and 2, and wash with physiological saline.

[0067] 2. Add 1 ml of sodium alginate solution with a concentration of 15 g / L to it to make a suspension.

[0068] 3. Use an electrostatic droplet generator to spray the suspension into 100ml of calcium chloride solution with a concentration of 100mmol / L to obtain a cell-containing calcium alginate bead with a diameter in the range of 100-500μm Precipitate, and remove the supernatant after the precipitation is complete.

[0069] 4. Add calcium alginate beads to 50ml of polylysine solution with a concentration of 0.5g / L, mix well, and place at room temperature for 5-10 minutes. After the precipitation is complete, remove the supernatant.

[0070] 5. Add the precipitate obtained in step 4 into 60ml of sodium alginate solution with a concentration of 1.5...

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PUM

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Abstract

The invention also discloses an immune isolated cell medicine, its preparing process and its application in killing tumors, wherein the product comprises semipermeable-immunological isolation film bundled active cells of the cutting immunological reactive substance, the reactive cells can excrete factors for suppressing tumor growth and / or eradicate tumor cells, its using process comprises injecting the medicament into tumor tissues or residual tumor tissues after surgery resection so as to eradicate tumor cells and / or suppress tumor tissue growth.

Description

field of invention [0001] The present invention relates to a novel immune isolation cell medicine, its preparation method and its application in killing and / or inhibiting tumors. Background technique [0002] At present, the cure rate of malignant tumors treated by three methods (surgical resection, chemotherapy, and radiotherapy) is very low. Surgery can only remove the tumor tissue that can be identified by the naked eye; radiotherapy or chemotherapy can only kill part of the tumor cells, and the remaining tumor cells become the root cause of tumor recurrence and metastasis; There are few areas and easy to produce drug resistance and other shortcomings. The treatment of malignant tumors is still a worldwide problem. [0003] Some protein factors can inhibit the growth of tumor tissue, but it is difficult to apply clinically. [0004] Studies have shown that some proteins and other factors can kill tumor cells and / or inhibit tumor tissue growth, such as: tumor necrosis f...

Claims

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Application Information

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IPC IPC(8): A61K45/00A61P35/00
Inventor 崔和厚
Owner 崔和厚
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