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Special combined mold for medicine screening and using method thereof

A technology of combining molds and molds, which is applied in the methods of supporting/immobilizing microorganisms, biochemical equipment and methods, bioreactor/fermenter combinations, etc.

Inactive Publication Date: 2021-10-22
中国医科大学
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Theoretically, the spindle-shaped complex organ with multi-branched tubular passages can be formed by the rotation of the mold, but the disadvantage is that the spindle forming height is different when the rotation speed of the mold is different. For different combination molds, the rotation speed is difficult to quantitatively limit. Therefore, it is difficult to determine the specific shape of the spindle
This combined mold is currently only used to shape the spindle-shaped organ precursor containing a set of branch channels, and it is easy to cause damage to the structure of the cell-containing matrix material when the inner mold is pulled out, and the cell activity is affected by the rotation
[0007] Invention patent (application number 201510419730.X) "A method and special mold for preparing tissue and organ precursors with multi-branched channels" proposes a special mold for preparing tissue and organ precursors with multi-branched channels, and its disadvantages The reason is that the intermediate operation steps are complicated, and the large and small pipes are easy to be misaligned
The hydrogel solution with a slightly higher viscosity is difficult to fill the entire cavity, and the mold in the branch cannot be disassembled, and it will damage the formed structure when it is taken out from the cell matrix layer

Method used

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  • Special combined mold for medicine screening and using method thereof
  • Special combined mold for medicine screening and using method thereof
  • Special combined mold for medicine screening and using method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0063] Example 1: Breast fabrication with multibranch vascular network and drug screening.

[0064] (1) Prepare a base mold, five vertical channel molds, horizontal channel molds, and orifice plate molds using photosensitive resin.

[0065] (2) Prepare 1% fibrinogen solution and thrombin solution (20 IU / ml).

[0066] (3) Coat all channel molds with a mixture of endothelial cells and fibrinogen, the cell density of this matrix solution is 5×10 7 cells / ml, add weight / volume (w / v) 1% heparin to the matrix solution, use thrombin solution (20 IU / ml) to polymerize fibrinogen for 2 min to obtain a tubular cell matrix layer.

[0067] (4) Assemble the various components of the combination mold: first, fix the orifice plate mold and the five vertical channel molds on the base mold through grooves, and then insert all the horizontal channel molds in (3) into the corresponding holes of the orifice plate mold in the hole. Pass the top to a cell density of 5 x 10 7 The mixed matrix solu...

Embodiment 2

[0070] Example 2: Brain production and drug screening with multi-branch blood vessels and neural networks.

[0071] (1) The base mold, five vertical channel molds, horizontal channel molds, and orifice plate molds were prepared using PTFE.

[0072] (2) 5% gelatin and 2% sodium alginate were blended and dissolved in PBS solution as a gelatin / sodium alginate matrix solution, in which endothelial cell growth factors (EGFs 0.5 ng / ml) were added. Prepare a 1% (w / v) calcium chloride solution.

[0073] (3) Coat all channel molds with a layer of adipose-derived stem cells (1×10 5 cells / ml) and endothelial cell growth factors (EGFs 0.5 ng / ml) in gelatin / sodium alginate matrix solution, and sodium alginate was cross-linked with 1% (w / v) calcium chloride solution to obtain a tubular endothelial cell layer .

[0074] (4) Assemble the various components of the combination mold: first, fix the orifice plate mold and the five vertical channel molds on the base mold through grooves, and th...

Embodiment 3

[0077] Example 3: Stomach manufacture and drug screening with multi-branched vessels and nerve conduit network.

[0078] (1) A base mold, five vertical channel molds, a horizontal channel mold, and an orifice plate mold were prepared using silicone rubber.

[0079] (2) Prepare 10% gelatin solution as matrix solution and 10% (w / v) transglutaminase (mTG, 100 U / ml) solution.

[0080] (3) Coat a layer of Schwann cells (1×10 3 a mixture of vascular epithelial cells and matrix solution on the other half of the channel mold, the density of cells in the matrix solution is 5×10 6 cells / ml, use transglutaminase to cross-link, and place at 37°C for 10 minutes to stabilize the structure of the gelatin-cell mixture and obtain a layer of tubular cell matrix layer. It is necessary to ensure that there is a certain amount of cell matrix layer on the surface of each tubular channel. thickness of.

[0081] (4) Assemble the various components of the special combination mold: first, fix the or...

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Abstract

The invention belongs to the technical field of medicine screening, and particularly relates to a special combined mold for medicine screening and a using method of the special combined mold. The complex organ with different cell matrix layers, which is provided with a middle large channel and a plurality of surrounding small channels, can be obtained. The combined mold includes: a base mold, an outer-layer pore plate mold, a middle-layer pore plate mold, an inner-layer frame mold, two central channel molds and a plurality of branch channel molds, wherein the inner-layer frame mold, the middle-layer pore plate mold and the outer-layer pore plate mold are sequentially nested from inside to outside to form a three-layer structure, and all layers do not make contact with one another and are embedded in the same base mold; the two central channel molds are positioned in the inner-layer frame mold and are embedded into the grooves of the base mold; a plurality of embedding holes are formed in the outer walls of the outer-layer pore plate mold, the middle-layer pore plate mold and the inner-layer frame mold, an embedding groove is further formed in the outer wall of the outer-layer pore plate mold, and the branch channel molds are inserted through the embedding holes or the embedding groove; finally, all the molds are combined to simulate a multi-channel complex organ.

Description

technical field [0001] The invention belongs to the technical field of drug screening, and in particular relates to a special combination mold for drug screening and its application method. Background technique [0002] Currently, chronic obstructive pulmonary disease (COPD), asthma, and pulmonary fibrosis (PF) have become lung diseases with high mortality rates. With the change of climate and environment, the lung diseases faced by human beings are showing an obvious increasing trend. For nearly half a century, with the continuous deepening of people's basic theoretical research on the lung and the positive progress made in research on the pathogenesis of some lung diseases, oxygen therapy, surgical treatment and rehabilitation therapy for lung diseases have made great progress. . Nevertheless, lung transplantation remains the mainstay of treatment for advanced lung disease. However, due to the lack of lung organ sources, many patients cannot get effective treatment at a...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12M3/00C12M1/34
CPCC12M21/08C12M23/20C12M23/44C12M25/14C12M29/12C12M41/46
Inventor 王小红徐宇坤宋达斌
Owner 中国医科大学