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Acellular matrix of human heart valve tissue and preparation and application of acellular matrix

A decellularized matrix and heart valve technology, applied in the field of biomedicine, can solve the problems of shortened time-consuming, long time-consuming, and easy depletion of cells, so as to save time, promote cell adhesion, and improve heart function.

Inactive Publication Date: 2015-05-06
RENJI HOSPITAL AFFILIATED TO SHANGHAI JIAO TONG UNIV SCHOOL OF MEDICINE
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  • Application Information

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Problems solved by technology

[0006] The invention creates a sheet-like decellularization method, which can solve the problem that the traditional substantive tissue decellularization method takes a long time and it is difficult to remove the cells in the middle part. After cutting into a membrane, the thickness of the membrane is close to that of the cells diameter, so most of the cells in the tissue can be exposed to the decellularization solution, it is easier to decellularize the cells, and the time-consuming can be greatly shortened

Method used

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  • Acellular matrix of human heart valve tissue and preparation and application of acellular matrix
  • Acellular matrix of human heart valve tissue and preparation and application of acellular matrix
  • Acellular matrix of human heart valve tissue and preparation and application of acellular matrix

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Embodiment

[0037] Repair of infarcted myocardium using human heart valve acellular matrix to construct myocardial patch:

[0038] 1. Preparation method of human heart valve tissue decellularized matrix:

[0039] Discarded human heart valve tissue from valve replacement patients (eg, figure 1 Shown in A) were embedded in the embedding agent O.C.T (Sakura Finetek, 4583, Japan), frozen at -20°C, and cut into thin slices with a thickness of 50 μm and thin slices with a thickness of 100 μm with a cryostat (Leica). The obtained flakes are subjected to decellularization treatment, specifically: the gained flakes are transferred into a 50ml centrifuge tube, adding mass fraction is 1% SDS (Sigma, L5750, U.S., the preparation method is to get 1g SDS, dissolve in 99g double-distilled water Autoclave for future use), placed in a shaker at a temperature of 37°C and a rotation speed of 180r / min for digestion for 12h. Wash with double distilled water several times to obtain digested human heart valve...

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Abstract

The invention provides an acellular matrix of a human heart valve tissue and a preparation and an application of the acellular matrix. The preparation method of the acellular matrix of the human heart valve tissue is characterized by comprising the following specific steps: cutting the human heart valve tissue into sheets which are 50-100microns in thickness; carrying out acellular disposal on the obtained sheets to obtain the acellular matrix of a heart valve; and freeze-drying and sterilizing for later use. The obtained human tissue is firstly cut into the sheets and then is subjected to decellularizing operation; the decellularizing time is greatly shortened when the decellularizing thoroughness is ensured, so that a lot of time is shortened; and the possibility of commercialization in future is ensured. The acellular matrix provided by the invention also proves that the acellular matrix of the human heart valve tissue has the effects of promoting stem cell adhesion, proliferation and myocardial direction differentiation in in-vitro cell culture, and is transplanted to the surface of the myocardial infarction heart, so that the heart function can be effectively improved. Therefore, the acellular matrix can also be applied to stem cell therapy of the heart disease as a new in-vitro cell culture medium and a transplant.

Description

technical field [0001] The invention relates to the preparation of cardiac tissue decellularized matrix and its basic research and application in cell culture and tissue engineering, and belongs to the fields of biomedicine, tissue engineering, cell biology, stem cell biology and heart and great vessel surgery. Background technique [0002] Cardiovascular disease is a global health problem. About one-third of the world's people die from cardiovascular disease. Myocardial infarction and congestive heart failure are the main causes of death from cardiovascular diseases, and myocardial infarction is an important cause of congestive heart failure. At present, the treatment of myocardial infarction is mainly to restore the blood flow of myocardial tissue around the ischemic area. Traditional methods such as drug heart protection, coronary artery stent implantation and bypass surgery are generally adopted, but these treatment measures can only protect the remaining myocardium. I...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61L27/36A61L27/38C12N5/071
Inventor 陈瑶万龙李军薛松
Owner RENJI HOSPITAL AFFILIATED TO SHANGHAI JIAO TONG UNIV SCHOOL OF MEDICINE
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