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Cardiac muscle tissue engineering product and its preparation method and use

A tissue engineering and product technology, applied in the field of myocardial tissue engineering products and their construction, can solve problems that have not yet been seen, and achieve the effect that is conducive to transportation

Active Publication Date: 2015-09-09
北京帝康医药投资管理有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, at present, the research on omentum acellular matrix has just started, and there is no report on the construction of engineered myocardial tissue using omentum acellular matrix as a scaffold material.

Method used

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  • Cardiac muscle tissue engineering product and its preparation method and use
  • Cardiac muscle tissue engineering product and its preparation method and use
  • Cardiac muscle tissue engineering product and its preparation method and use

Examples

Experimental program
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Effect test

preparation example Construction

[0035] According to one aspect of the present invention, a preparation method of a myocardial tissue engineering product is provided, which includes:

[0036] (1) preparing the omentum decellularized matrix scaffold material, which includes the step of soaking the omentum in a decellularization solution containing sodium lauryl sulfate, DNase and trypsin for decellularization; and

[0037] (2) Construct and culture cardiomyocyte-omentum acellular matrix complex.

[0038] In some embodiments, step (1) further includes a degreasing treatment step. In an exemplary embodiment, the step of defatting treatment is performed prior to the step of decellularization treatment. In some preferred embodiments, the step of degreasing treatment is carried out using a mixed solution of methanol, chloroform and ether at a volume ratio of 1:1:0.2-1. In certain more preferred embodiments, the volume ratio of methanol, chloroform and diethyl ether is 1:1:0.4-0.7. In the most preferred embodimen...

Embodiment 1

[0064] Embodiment 1 prepares cardiomyocyte suspension

[0065] Cut myocardial tissue into 1mm under sterile conditions 3 size. After washing with PBS, add 0.25% trypsin solution, pipette gently, absorb the supernatant and add to the whole culture medium containing 10% serum to terminate the digestion. Repeat the above steps until the digestion is complete. Use a 200-mesh sieve to collect the digestion solution, centrifuge to collect the cells, resuspend the cells in the culture medium, inoculate in a 10cm cell culture dish, and place in CO 2 Constant temperature incubator. Differentially adhere to the wall for 1 hour to remove fibroblasts, collect unattached cells, and divide 2×10 6 Inoculate a 10cm cell culture dish at the seeding density of cells / dish and place in CO 2 cultured in an incubator.

[0066] Subsequently, the cultured cardiomyocytes were digested by trypsin, the cells were resuspended in DMEM medium, and the cell density was prepared to be 5×10 6 / mL of ca...

Embodiment 2

[0067] Example 2 Preparation of Greater Omentum Acellular Matrix

[0068] Fresh minipig peritoneal omentum tissues (purchased from slaughterhouses) were washed 3 times in PBS. The cleaned tissues were soaked in 2L of degreasing solution for 24 hours by shaking, and the shaking frequency was 200r / min. Subsequently, the fat-removed omentum tissue was washed three times in PBS, and soaked in 1L of decellularized solution for 18 hours with shaking at a frequency of 150r / min. The omentum tissue treated with the decellularized solution was freeze-dried, sterilized by irradiation, and stored at room temperature.

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PUM

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Abstract

The invention discloses a cardiac muscle tissue engineering product and its preparation method and use. The preparation method comprises 1, omentum majus decellularized matrix scaffold material preparation: immersing omentum majus in a decellularizing liquid containing lauryl sodium sulfate, DNA enzyme and trypsin so that decellularizing treatment is realized and 2, myocardial cell-omentum majus decellularized matrix complex construction and culture. The cardiac muscle tissue engineering product comprises the myocardial cell and omentum majus decellularized matrix scaffold material. The omentum majus decellularized matrix has a good 3D netted space structure and is rich in a growth factor so that myocardial cell growth and nutrient and metabolite transport are realized. The cardiac muscle tissue engineering product has an important meaning for cardiac muscle damage restoration based on the cardiac muscle tissue engineering product and cardiac muscle treatment drug screening utilizing the cardiac muscle tissue engineering product as a model.

Description

technical field [0001] The invention belongs to the technical field of tissue engineering, and in particular relates to a myocardial tissue engineering product and a construction method thereof using the omentum acellular matrix as a scaffold material. Background technique [0002] Myocardial tissue engineering is a combination of the principles and technologies of cell biology, biomaterials and engineering, combining seed cells with the ability to form myocardial tissue with bioscaffold materials with good biocompatibility, degradability and certain biomechanical properties. Compounding, in vitro construction of engineered myocardial tissue capable of myocardial transplantation and repair of myocardial injury, the main research content involves three aspects: seed cells, scaffold materials, and engineered myocardial tissue construction. Among them, the selection of biological scaffold materials is the key to the construction of engineered myocardial tissue. [0003] Tissue...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61L27/38A61L27/50A61L27/54
Inventor 饶义伟蓝德宾
Owner 北京帝康医药投资管理有限公司
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