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Cell-microspheres with eluted superparamagnetic gel coating and preparation method and application thereof

A gel microsphere and superparamagnetic technology, which is applied in the field of cell-microsphere and its preparation, can solve the problems that cannot be repaired by clinical implantation, cannot be removed by magnetic materials, and can separate different cells, so as to avoid the residual magnetic material. Effect

Active Publication Date: 2022-07-08
SICHUAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The purpose of the present invention is to overcome the deficiencies in the prior art, to provide a cell-microsphere with an eluted superparamagnetic gel coating and its preparation method and application, to solve the problem that the existing microcarriers cannot Realize the removal of magnetic materials and the separation of different cells in the co-culture system, which cannot be used for clinical implant repair and other issues

Method used

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  • Cell-microspheres with eluted superparamagnetic gel coating and preparation method and application thereof
  • Cell-microspheres with eluted superparamagnetic gel coating and preparation method and application thereof
  • Cell-microspheres with eluted superparamagnetic gel coating and preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0064] In this example, the steps of preparing collagen gel microspheres loaded with bone marrow mesenchymal stem cells are as follows:

[0065] (1) Take a collagen solution with a concentration of 15 mg / mL, adjust the pH to 7.4 with 1 mol / L sodium hydroxide solution in an ice-water bath, and then dilute the collagen solution to a collagen concentration of 10 mg / mL; Bone marrow mesenchymal stem cells were counted and prepared into a cell suspension with α-MEM medium, added to the collagen solution and dispersed uniformly to obtain a cell-collagen mixture. In the cell-collagen mixture, the final concentration of collagen was 6.5 mg / mL, The cell density is 5×10 6 pcs / mL.

[0066](2) Take 70 mL of methyl silicone oil with a viscosity of 50 mPa·s, add 0.05% (v / v) Span-80 to form an oil phase, stir with a magnetic stirrer at a speed of 500 rpm, and under ice-water bath conditions, cells- The collagen mixture (water phase) was added dropwise to the oil phase, and stirring was cont...

Embodiment 2

[0068] In this example, the preparation of collagen gel microspheres loaded with fluorescent particles is as follows:

[0069] (1) Take a collagen solution with a concentration of 15 mg / mL, adjust the pH to 7.4 with 1 mol / L sodium hydroxide solution in an ice-water bath, and then dilute the collagen solution to a collagen concentration of 6 mg / mL; take the red fluorescent particles and Green fluorescent particles (v / v: 1 / 50) were uniformly dispersed in the collagen solution to obtain a fluorescent particle-collagen mixture.

[0070] (2) Take 60 mL of methyl silicone oil with a viscosity of 50 mPa·s, stir with a magnetic stirrer at a rotational speed of 550 rpm, and add the fluorescent particle-collagen mixture (water phase) dropwise to the oil phase under ice-water bath conditions, and continue After stirring for 30 minutes, the water phase-oil phase mixed system was heated to 37°C, and the stirring was continued for 20 minutes. After the stirring was completed, the obtained m...

Embodiment 3

[0072] In this example, the steps of preparing cell-microspheres with eluted superparamagnetic gel coating are as follows:

[0073] (1) Take the bone marrow mesenchymal stem cells-collagen gel microspheres prepared in Example 1, place them in a centrifuge tube, wash 3 times with a sodium chloride solution, and then soak them in a sodium chloride solution containing calcium chloride 3 minutes, drain the liquid and set aside.

[0074] (2) Dilute the 6% sodium alginate solution by one time with a solution containing 4 mg / mL superparamagnetic particles with a diameter of 1 μm, and mix evenly to obtain a coating material for use.

[0075] (3) adding the coating material prepared in step (2) dropwise to the bone marrow mesenchymal stem cells-collagen gel microspheres treated in step (1), mixing evenly, and adding dropwise to the surface of the gel microspheres The coating material reacts with calcium ions on the surface of the gel microspheres to gel to form a gel coating. The exce...

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Abstract

The present invention provides a cell-microsphere with an eluted superparamagnetic gel coating and a preparation method thereof. The cell-microsphere comprises a gel microsphere and a super-paramagnetic sphere coating the gel microsphere. Magnetic gel coating, the gel microspheres contain cells, the superparamagnetic gel coating contains superparamagnetic particles, the superparamagnetic gel coating can be removed by elution, and the superparamagnetic gel The gel coat and gel beads cannot be removed by simultaneous elution. The present invention also provides the application of the above-mentioned cell-microspheres in the co-culture of cells containing cell-gel microspheres for clinical implantation and repair and the separation in the later stage of the co-culture. The invention can solve the problems that the existing microcarrier cannot realize the removal of magnetic material and the separation of different cells in the co-culture system after the co-culture, and cannot be used for clinical implantation and repair.

Description

technical field [0001] The invention belongs to the technical field of biomedical macromolecular materials, and relates to a cell-microsphere with eluted superparamagnetic gel coating and a preparation method and application thereof. Background technique [0002] The existing magnetic microcarriers are obtained by treating the surface of magnetic materials, wrapping polymer materials, and binding antigens, antibodies, nucleic acids and other biological macromolecules. The magnetic materials are located inside the microcarriers, and most of the magnetic microcarriers are later-planted cells. , cells are grown on the surface of the microcarriers. For example, the antibody-coated magnetic beads are fully interacted with the cells to be separated, and the antibody magnetic beads are used to combine with the corresponding cells into the form of cell-antibody-magnetic bead complexes, using their movement in the magnetic field to interact with other cells. differences, cells can b...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/00C12N5/0775
CPCC12N5/0075C12N5/0663C12N2513/00C12N2531/00
Inventor 樊渝江刘钧王启光孙勇林海
Owner SICHUAN UNIV
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