Preparation method and application of fibronectin-functional cell composite type microencapsulation transplant

A technology of fibronectin and microencapsulation, applied in the field of biomedicine, can solve the problems of short survival period of functional cells, poor long-term treatment effect, shortage of active functional cells, etc., shorten the time of cell confluence and eliminate the risk of heterogeneous infection , Promote the effect of body cell and tissue regeneration

Inactive Publication Date: 2017-07-21
王斌
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Problems solved by technology

[0007] The purpose of the present invention is based on the above-mentioned state of the art, by establishing a method for preparing a composite microencapsulated graft of fibronectin and functional cells, to solve the current clinical application of cell transplantation in the treatment of major diseases such as cardiovascular disease and diabetes.

Method used

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Examples

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Example Embodiment

[0038] Example 1 Preparation method of "Fibronectin and bovine cardiomyocyte composite microencapsulated graft":

[0039] 1. Selection of bovine cardiomyocyte donor: SPF newborn bovine is selected as the donor of cardiomyocytes.

[0040] 2. Isolation, purification and identification of bovine cardiomyocytes: Myocardial tissue was dissected from SPF newborn cattle and placed in a sterile dish, added with an appropriate amount of pre-cooled PBS, washed away blood clots and blood cells, removed blood vessels, fat, and connective tissue, and then Rinse 2-3 times with pre-cooled PBS, transfer to a scintillation vial filled with pre-cooled PBS, and cut the myocardial tissue into 1mm 2 Uniform pieces of size. Add 0.12% type I collagenase, add magnets to the scintillation vial, and digest for 10 min in a 37 ℃ water bath at 250 r / min. Discard the supernatant after the first digestion, add new digestive enzymes to continue the digestion for 10 minutes, and repeat the digestion 4-5 times unt...

Example Embodiment

[0043] Example 2 Preparation method of "fibronectin and porcine islet cell composite microencapsulated transplant":

[0044] 1. Isolation of porcine pancreatic islets: dissect the pancreas from SPF pigs under clean conditions, place the pancreas on a clean workbench, quickly remove the peripancreatic fat, blood vessels and pancreatic capsule, retain the proper capsule, rinse with UW solution and weigh with a balance . Transect the pancreas from the junction of the head and tail of the pancreas, find the main pancreatic duct, insert the plastic tube, and perfuse the collagenase solution (1.5 g / L, containing DNase 0.3 g / L, containing 2 times the weight of the pancreas) of collagenase solution prepared by Hanks solution. L). Digestion in 37℃ water bath for 30 minutes, pH is about 7.5 during digestion, samples are taken every 4 minutes from the beginning of digestion 20 minutes, dithizone staining microscopy, when pancreas tissue is digested and broken into fine sand, microscopic ex...

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Abstract

The invention relates to a preparation method and application of a fibronectin-functional cell composite type microencapsulation transplant. The method includes: selecting an adult or an SPF mammal as a functional cell donor, separating cells with specific functions from the adult or the SPF mammal, and purifying the separated cells; using an FN serum-free culture medium to perform in-vitro culture on the purified functional cells so as to obtain a large amount of high-quality active functional cells through amplification; using the microencapsulation immune isolation technology to jointly embed fibronectin and the functional cells into an APA micro-capsule according to a certain proportion so as to form the fibronectin-functional cell composite type microencapsulation transplant. The preparation method has the advantages that technical difficulties that high-quality active functional cells are insufficient, allosome or xenogeneic immunological rejection exists, the surviving period of transplanted functional cells in a receptor is short, the long-term treatment effect is poor, and the like in the current clinical application using cell transplanting to treat serious diseases such cardiovascular disease and diabetes can be solved, and a new technical manner and method using the cell transplanting to treat various serious diseases is provided.

Description

technical field [0001] The invention relates to the field of biomedicine, in particular to a preparation method and application of a composite microencapsulated graft of fibronectin and functional cells. Background technique [0002] With the development of social economy and the change of people's lifestyle, more and more diseases affect human health, among which cardiovascular disease and diabetes have a large population base, high morbidity and high mortality have become global threats to human health. Major diseases. [0003] Cardiovascular disease (CVD) has always been a disease with the highest morbidity and mortality worldwide. Recently, the latest data from the World Health Organization (WHO) shows that cardiovascular disease has become the leading cause of death in the world. In 2004, 17.1 million people died of CVD, accounting for 29% of global deaths, of which about 7.2 million died of coronary heart disease, 570 Wan died of a stroke. It is estimated that by 20...

Claims

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Application Information

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IPC IPC(8): A61K35/39A61K35/34A61K35/44A61K38/39A61K47/34A61K47/36A61K9/50A61P9/00A61P3/10C12N5/071C12N5/077
Inventor 王斌桑喜成张世阳许义辉贾秀秀杨敏祺
Owner 王斌
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