Ultrathin carrier cell sheet and preparation method thereof

A cell sheet and carrier technology, applied in prosthetics, coatings, medical science, etc., can solve the problems of easy damage, poor mechanical properties of cell sheets, and artificial modification of cell sheets that cannot be loaded with drugs, so as to achieve precise shape control and flexibility Ease of machinability, effect of material reduction

Active Publication Date: 2017-07-07
ZHEJIANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the cell sheets constructed by these methods connect the cells to each other through the structure between cells and the extracellular matrix (extracellular mix, ECM) secreted by cells, so the mechanical properties of this cell sheet are poor and easy to break
And, because there is no additional material structure, such cell sheets cannot be loaded with drugs or further artificially modified

Method used

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  • Ultrathin carrier cell sheet and preparation method thereof
  • Ultrathin carrier cell sheet and preparation method thereof
  • Ultrathin carrier cell sheet and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0040] Prepare an aqueous solution of gelatin with a mass fraction of 1%, wait for it to dissolve at 55°C, apply it evenly on the surface of the cover glass, put it in a vacuum drying oven, and vacuum at room temperature for 6 hours to remove the solvent.

[0041]Take out the glass slide, immerse in polystyrene sulfonate aqueous solution for 5 minutes, then immerse in water for 3 minutes, then immerse in polyallylamine hydrochloride solution for 5 minutes, and then immerse in water for 3 minutes. Carry out 15 cycles in this order, dry naturally, and sterilize with ethylene oxide gas. Wherein polystyrene sulfonate, polypropylene amine hydrochloride aqueous solution, concentration 2g / L, add 0.15M sodium chloride, adjust pH value to 3.

[0042] Place the glass slides obtained in the previous step in a petri dish, and add the rat-derived bone marrow mesenchymal stem cell suspension (10 4 ~10 6 cells / ml) were submerged on the surface, and cultured in a conventional cell culture i...

Embodiment 2

[0044] Prepare an aqueous solution of gelatin with a mass fraction of 15%, wait for it to dissolve at 55°C, spin-coat it on the surface of a glass slide with a coater, put it in a vacuum drying oven, and vacuum it at room temperature for 6 hours to remove the solvent.

[0045] Take out the glass slide, immerse in hyaluronic acid aqueous solution for 10 minutes, then immerse in water for 3 minutes, then immerse in lysine aqueous solution for 10 minutes, and then immerse in water for 3 minutes. Carry out 40 cycles in this order, dry naturally, and sterilize with ethylene oxide gas. Wherein hyaluronic acid, polylysine aqueous solution, concentration 1g / L, add 0.15M sodium chloride, adjust pH value to be 3.

[0046] The glass slide obtained in the previous step was placed in a petri dish, and the corneal epithelial cell suspension (10 4 ~10 6 cells / ml) were submerged on the surface, and cultured in a conventional cell culture incubator for 24 hours. The cell sheet is detached f...

Embodiment 3

[0048] Prepare a gelatin ethanol solution with a mass fraction of 5%, dissolve it at 55°C, apply it evenly on the surface of a glass slide, put it in a vacuum drying oven, and vacuum at room temperature for 6 hours to remove the solvent.

[0049] Take out the glass slide, immerse in polystyrene sulfonate aqueous solution for 20 minutes, then immerse in water for 5 minutes, then immerse in polyallylamine hydrochloride aqueous solution for 20 minutes, and then immerse in water for 5 minutes. Carry out 40 cycles in this order, dry naturally, and sterilize with ethylene oxide gas. Wherein polystyrene sulfonate, polypropylene amine hydrochloride aqueous solution, concentration 2g / L, add 0.15M sodium chloride, adjust pH value to 3.

[0050] The glass slide obtained in the previous step was placed in a petri dish, and the corneal epithelial cell suspension (10 4 ~10 6 cells / ml) were submerged on the surface, and cultured in a conventional cell culture incubator for 24 hours. The c...

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Abstract

The invention discloses an ultrathin carrier cell sheet and a preparation method thereof. The preparation method comprises the following steps of (1) spreading a gel material solution onto a substrate, and condensing into a solid state; (2) generating an ion polymer layer which has opposite polarity with the gel material on the substrate spread with the gel material, generating an ion polymer layer which has opposite polarity with the upper ion polymer layer, on the upper ion polymer layer, and repeating the operation, so as to form a layered self-assembly film; (3) planting a target cell into the substrate obtained in step (2), and conventionally culturing; (4) in the culture process, enabling the layered self-assembled film loaded with the cell to disengage from the substrate, so as to form the cell sheet. Compared with the traditional preparation method of the cell sheet, the preparation method has the advantages that the character control or decomposing step of a supporter material is not needed, the mechanical strength of the cell sheet is obviously improved, and macromolecular medicines, such as protein, polypeptide, antibody and nucleic acid, can be loaded.

Description

technical field [0001] The invention belongs to the field of tissue engineering, in particular to an ultra-thin carrier cell sheet and a preparation method thereof. Background technique [0002] Cell transplantation is widely used in the field of tissue engineering for cornea, myocardial reconstruction, and tissue repair. However, the traditional method of injecting cell suspension cannot ensure the effective residence of cells in the damaged area, nor can it control the shape and size of the repaired area. Moreover, since the cell suspension needs to be treated with trypsin or chemicals during the collection process, it can be stripped and recovered from the surface of the culture container, which will cause cell damage and damage the original function. Therefore, cells are seeded onto biodegradable scaffold materials such as collagen gel and fibrin gel, and then transplanted together with the scaffold material. This cell-containing scaffold has tunable mechanical propert...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61L27/44A61L27/38A61L27/18A61L27/20A61L27/16A61L27/22A61L27/54A61L27/50C08J7/04C08J7/06
CPCA61L27/3834A61L27/3891A61L27/50A61L27/54A61L2300/232A61L2300/252A61L2300/256C08J7/042C08J7/06C08J2389/00C08J2405/08C08J2425/06
Inventor 王本贺川江叶婷婷
Owner ZHEJIANG UNIV
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