Construction method of tissue engineered tissue based on artificial cell

An artificial cell and tissue engineering technology, which is applied in the field of tissue engineering tissue construction, can solve the problems of low connectivity, reduce the porosity and connectivity of scaffold materials, reduce the impact, realize large-scale production, and improve permeability Good results

Active Publication Date: 2013-10-16
南通美韦德生命科学有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0005] It can be seen from the above that researchers are constantly exploring new ways of tissue construction while constructing tissue, and have achieved some good results, but there are still some problems between different tissue construction methods, for example, the scale of in vivo construction, The low connectivity rate of microsphere fusion construction, the tissue size obtained by scaffold-free culture, the influence of hydroge

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] Step 100: Take two bottles of confluent normal-growing chondrocytes, remove the old medium, wash the cells twice with D-PBS, 2-3 mL each time, and remove the washing solution. Add 500 μL of trypsin to make it completely cover the cell layer, and place in CO 2 Incubate for about 1 min in the incubator. When the cells are about to separate and become round particles, tap the bottom of the culture bottle to make the cells fall off, add 4mL of complete culture medium (DMEM medium containing 10% fetal bovine serum, 1% double antibody, the above are all volume fractions) and mix well Form a cell suspension and terminate the digestion with trypsin. Count, the total number of cells is about 1×10 7 Transfer the cell suspension to a sterile 15mL centrifuge tube and centrifuge at 1000rpm for 5min. Remove the supernatant, add 0.5mL fresh complete medium and pipette to resuspend the cells.

[0028] Step 10. Add the above cell suspension to 2mL of 1.5% Alg (sodium alginate soluti...

Embodiment 2

[0035] The difference between this embodiment and the first embodiment is that the cells are liver cells, and the hydrogel material is silk fibroin. Using an ultrasonic cell breaker, the silk fibroin solution is ultrasonicated for 1 min at a power of 200W, and the artificial cells and the silk fibroin solution are mixed uniformly at a volume ratio of 1:2, and the obtained mixture is allowed to stand at room temperature for 5 min to obtain artificial cells. -Silk fibroin gel complex.

Embodiment 3

[0037]This example is basically the same as Example 1, except that the culture condition is a dynamic shaker.

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PUM

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Abstract

The invention provides a construction method of a tissue engineered tissue based on an artificial cell. The method comprises the following steps of: floating a cell suspension in a filtered and sterilized sodium alginate solution, and dropping the cell suspension into a calcium chloride solution through a dropping liquid generation device to form a calcium alginate bead; placing the bead into a membrane material contained solution, and coating a membrane on the surface of the bead to form a microcapsule, wherein the bead is cleaned by using normal saline; then, neutralizing superfluous positive charges on the surface of the microcapsule; liquefying the calcium alginate bead in the microcapsule by using a sodium citrate solution to form the artificial cell; next, curing and forming a mixture of the artificial cell and an aquogel to finally form an artificial cell and aquogel complex; and placing the artificial cell and aquogel complex under a static or oscillatory or microgravity condition to culture to construct the tissue engineered tissue. By using the construction method, the requirement of the traditional support for high porosity and high connectivity rate can be reduced; and the construction method is simple and beneficial to the realization of large-scale production for in-vitro construction.

Description

【Technical field】 [0001] The invention relates to the technical field of tissue engineering, in particular to a method for constructing tissue engineered tissue based on artificial cells. 【Background technique】 [0002] Tissue engineering is currently a hot research field at home and abroad, and how to successfully construct cell-scaffold complexes to form tissues is a key issue in tissue engineering research. [0003] The current basic idea of ​​tissue engineering tissue construction is to prepare scaffold materials first, then transplant seed cells onto the scaffold materials, and after a certain period of culture, construct tissues and organs in vitro or in vivo. The research on scaffold materials is very extensive. Various synthetic materials, natural biomaterials and composite materials have been widely used, and through rapid prototyping technology, tissue engineering scaffold materials that fully match the three-dimensional structure of defective tissues have been man...

Claims

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Application Information

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IPC IPC(8): A61L27/38A61L27/20A61L27/52A61L27/50
Inventor 刘源岗王士斌何鹏林琴汤晓琳陈爱政
Owner 南通美韦德生命科学有限公司
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