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A kind of preparation method of sandwich-like multicellular sheet

A multi-cell, sandwich technology, applied in the field of biomedical tissue engineering, can solve the problems of internal cell necrosis and difficulty in obtaining seed cells, and achieve high biomimetic, simple and easy operation methods, and good biocompatibility.

Inactive Publication Date: 2016-10-26
THE SECOND PEOPLES HOSPITAL OF SHENZHEN
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Like other tissue engineering methods, when cell sheet technology is used for tissue reconstruction, it is difficult to obtain seed cells; when the superimposed cell sheet exceeds a certain thickness, it often leads to necrosis of internal cells; the application of cell sheet technology to construct large functional Sexual tissue structure remains a major challenge for cell sheet technology

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] Dissolve 0.12g chitosan in 4ml 50% trifluoroethanol solvent, gradually add 0.48g collagen during magnetic stirring, stir for 2h until completely dissolved, and obtain 4ml 15% mixed spinning solution. Inhale the mixture into the syringe, control the advance speed of the micro-injection pump to 0.5ml / h, select a No. 9 stainless steel needle and connect it to a 9.5kv high voltage, and use a grounded aluminum foil to receive the fiber at a distance of 9cm from the needle tip. After 8 hours, a disordered nanofiber film with an average diameter of about 170 nm was formed on the aluminum foil, which had poor mechanical strength and was brittle and easily broken. The prepared electrospun biofilm was sterilized by UV irradiation for 15 min. Place the sterilized biofilm on the adipose stem cell sheet that has grown and merged on the surface of the temperature-sensitive culture dish, lower the temperature of the culture surface to 25°C, peel off the entire cell sheet from the edge...

Embodiment 2

[0030] Dissolve 0.18g chitosan in 4ml 50% trifluoroethanol solvent, gradually add 0.42g collagen during magnetic stirring, stir for 2h until completely dissolved, and obtain 4ml 15% mixed spinning solution. Inhale the mixture into the syringe, control the advance speed of the micro-injection pump to 0.5ml / h, select a No. 8 stainless steel needle and connect it to a 10kv high voltage, and use a grounded aluminum foil to receive the fiber filament at a distance of 10cm from the needle tip. After 8 hours, a disordered nanofiber film with an average diameter of about 200 nm was formed on the aluminum foil, which had poor mechanical strength and was brittle and easily broken. The prepared electrospun biofilm was sterilized by UV irradiation for 15 min. Place the sterilized biofilm on the adipose stem cell sheet that has grown and merged on the surface of the temperature-sensitive culture dish, lower the temperature of the culture surface to 25°C, peel off the entire cell sheet from...

Embodiment 3

[0032] Dissolve 0.3g of chitosan in 4ml of 50% trifluoroethanol solvent, gradually add 0.3g of collagen during magnetic stirring, stir for 3 hours until completely dissolved, and obtain 4ml of 15% mixed spinning solution. Inhale the mixture into the syringe, control the advance speed of the micro-injection pump to 0.8ml / h, select a No. 7 stainless steel needle and connect it to a high voltage of 11kv, and use a grounded aluminum foil to receive the fiber filament at a distance of 12cm from the needle tip. After 5 hours, a disordered nanofiber film with an average diameter of about 300 nm was formed on the aluminum foil. The prepared electrospun biofilm was sterilized by UV irradiation for 15 min. Place the sterilized biofilm on the adipose stem cell sheet that has grown and merged on the surface of the temperature-sensitive culture dish, lower the temperature of the culture surface to 22°C, peel off the entire cell sheet from the edge after 20 minutes, and then turn it upside ...

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Abstract

The invention discloses a preparation method of a sandwich-shaped multicellular lamella. The preparation method comprises the following steps: an electrospinning liquid preparation step: adding chitosan and collagen in a trifluoroethanol solution, and stirring to obtain an electrospinning liquid, wherein the mass ratio of the chitosan to the collagen ranges from (1: 4) to (4: 1); an electrostatic spinning step: carrying out high-voltage electrostatic spinning on the electrospinning liquid to obtain a chitosan-collagen electrospinning biomembrane; and a cell-membrane-cell multicellular lamella construction step: respectively adhering two cell lamellas to the two surfaces of the chitosan-collagen electrospinning biomembrane to grow, so as to obtain a cell-membrane-cell multicellular lamella. According to the preparation method disclosed by the invention, the preparation process is simple and controllable; with the electrospinning biomembrane as a support membrane, as the two cell lamellas are adhered to the biomembrane to grow, the proliferation space of cells is enlarged, circulation of nutrients between the two cell lamellas is improved, and the integrity of the cell lamellas in the transfer process is also ensured, thus a simple, convenient and easy operation method is available for clinical multicellular lamella transplantation therapy.

Description

technical field [0001] The invention relates to the field of biomedical tissue engineering, in particular to a method for preparing a multi-cell sheet. Background technique [0002] With the development of life sciences, it has become a hot spot in modern medicine to replace and regenerate tissue defects caused by injury and disease by tissue engineering. The basic elements of tissue engineering technology include tissue cells with good response characteristics, supporting extracellular matrix and bioactive factors that promote cell and tissue regeneration. The most important elements of tissue engineering are seed cells and scaffolds. The key technology is to inoculate seed cells with plastic degradable biomaterials and implant them into the body to grow to form the desired tissue. There are generally two ways to repair defects in traditional tissue engineering: one is to directly inject or place cells or cell clusters in the defect; the other is to inoculate cells on biod...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61L27/20A61L27/24A61L27/50C12N5/0775
Inventor 朱艳霞余小平
Owner THE SECOND PEOPLES HOSPITAL OF SHENZHEN
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