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Method for preparing microcapsulized functional cell

A microencapsulation and cell technology, applied in biochemical equipment and methods, biological material sampling methods, microcapsules, etc., can solve the problems of unsatisfactory purity and activity of recombinant cytokines, short biological half-life, toxic and side effects, etc. , to achieve the effect of good permeability and biocompatibility, small effect on activity and low cost

Inactive Publication Date: 2003-08-27
SECOND AFFILIATED HOSPITAL ZHEJIANG UNIV COLLEGE OF MEDICINE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, due to the short biological half-life of cytokines in the body, continuous high-dose intravenous administration or intermittent subcutaneous injection is necessary to ensure effective therapeutic concentrations
However, large doses of medication can often cause serious side effects; at the same time, the purity and activity of recombinant cytokines are not satisfactory

Method used

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  • Method for preparing microcapsulized functional cell
  • Method for preparing microcapsulized functional cell
  • Method for preparing microcapsulized functional cell

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0020] see figure 1 , a structure of a multi-channel microcapsule generating device includes: a piston type syringe pump 1, connecting pipes 2 and 2-1, a pipe joint 3, a tube core 4, a gland 5, a sealing ring 6, a fixing ring 7, a jacket 8, Compressed gas generator 9, the connecting pipe 2 has multiple passages, one of which is connected to the piston type syringe pump 1, and the remaining passages are connected to the corresponding number of dies 4 through the pipe joint 3, and the other end of the die 4 is a microcapsule The outlet of the tube core 4 passes through the center of the gland 5, the sealing ring 6, and the fixing ring 7 in sequence, the sealing ring 6 is located between the gland 5 and the fixing ring 7, and the outer jacket 8 is set on the tube core 4, the sealing ring 6, and the fixing ring 7. Outside the circle 7, a cylinder is formed, the upper end of the cylinder is covered with a gland 5, and the lower end becomes the outlet of the microcapsule. There is a...

Embodiment 2

[0022] Microencapsulation of cells: Take CHO cells that are in good growth state and reach 90% confluence, wash twice with 0.25% trypsinized saline, centrifuge and count, take the filter-sterilized microencapsulation solution sodium alginate, and put Cells are scaled (1.5 x 10 9 L -1 Sodium alginate) was mixed in the sodium alginate solution, and mixed evenly by pipetting gently. Spray the cell suspension into a solution containing 4.9% BaCl using a multi-channel microcapsule generator 2 solution (pH7.4) in the receiving tank, after 10 min, washed 3 times with 1×PBS solution. The microcapsules were collected, and the living conditions of the cells in the capsules were dynamically observed under a microscope. Placed in DMEM containing 10% calf serum, cultured in an incubator (37°C, 5% CO 2 )spare.

[0023] The microcapsule encapsulation liquid used can also be polylysine, agarose, polyallylamine and the like.

[0024] Microcapsules with different diameters (φ0.35-0.75mm) ...

Embodiment 3

[0025] The mensuration of embodiment 3 sodium alginate microcapsule physical and chemical properties (one) experimental material

[0026] Instrument materials: laser scanning confocal microscope (LSCM 510); constant temperature oscillator TH2-c; isothiocyanate fluorescently labeled dextran (FITC-dextran, FD) (FD-40, FD-75 and FD-167) . FITC-labeled goat anti-mouse IgG (F-IgG), trypsin; DMEM medium, newborn bovine serum. CHO cells.

[0027] Experimental animals: BALB / c mice, 8-10 weeks old, weighing 20-25g. (2) Determination of physical and chemical properties of microcapsules 1. Permeability measurement and static observation of microcapsules

[0028] Suspend the various microcapsules prepared above in 0.5ml solution containing FD-40, FD-75 or FD-167 (both 100mg / L) and F-IgG solution (100mg / L), and place them at room temperature and away from light 24hr, and keep shaking, so that the fluorescein label can fully diffuse into the microcapsules. Adjust the laser scanning con...

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Abstract

The invention is a method of making microencapsulated functional cell. It mixes evenly the filtered and axenic bubble enveloping liquid and good cells in proportion, then places the mixture in the multichannel generating equipment, spray the cell suspended liquid to form bubble, and then processes and reserves. The generating equipment includes injection pump, connecting hose (2-1), union joint, hose core, gland, sealing ring, fixing ring, jacket and compressed gas generator.

Description

technical field [0001] The invention belongs to the technical field of biological treatment, and relates to a preparation technology of biological microcapsules, in particular to a preparation method of microencapsulated functional cells, which can prepare microencapsulated functional cells with high histocompatibility, permeability and firmness. Cells can be effectively applied to the biological treatment of tumors. technical background [0002] In recent years, as the fourth treatment mode of malignant tumors, biological therapy has been further developed, and its position in tumor treatment has become more and more important. Among them, the application of genetically engineered cytokines, tumor immunotherapy and tumor gene therapy are the most widely studied and applied, and have achieved certain results. However, due to the short biological half-life of cytokines in the body, continuous high-dose intravenous administration or intermittent subcutaneous injection is nece...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K9/50A61K45/00A61P35/00C12M1/00C12N5/00
CPCC12M33/06
Inventor 郑树孝作祥潘月龙董琦
Owner SECOND AFFILIATED HOSPITAL ZHEJIANG UNIV COLLEGE OF MEDICINE
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