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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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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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. The shortage of high-quality active functional cells, allogeneic or heterogeneous immune rejection, the short survival period of functional cells in the recipient after transplantation, and poor long-term therapeutic effect, etc., are three major technical problems for cell transplantation therapy. Disease provides a new technical means and method

Method used

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Examples

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Effect test

Embodiment 1

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

[0039] 1. Selection of bovine cardiomyocyte donors: SPF newborn cattle were selected as cardiomyocyte donors.

[0040] 2. Separation, purification and identification of bovine cardiomyocytes: Dissect myocardial tissue from SPF newborn cattle and place it in a sterile plate, add an appropriate amount of pre-cooled PBS, wash away blood clots and blood cells, remove blood vessels, fat, and connective tissue, and then Rinse 2-3 times with pre-cooled PBS, transfer to scintillation vials filled with pre-cooled PBS, cut myocardial tissue into 1mm 2 Evenly sized pieces. Add 0.12% type Ⅰ collagenase, add magnetic particles to the scintillation vial, and digest in a water bath at 37 °C at 250 r / min for 10 min. Discard the supernatant after the first digestion, add new digestive enzymes to continue digestion for 10 min, and repeat digestion 4-5 times until the tissue turns...

Embodiment 2

[0043] Example 2 Preparation method of "Fibronectin and Porcine Islet Cell Composite Microencapsulated Transplant":

[0044] 1. Porcine islet isolation: dissect the pancreas from SPF pigs under clean conditions, place the pancreas on an ultra-clean workbench, quickly remove peripancreatic fat, blood vessels and pancreatic capsule, retain the intrinsic capsule, wash with UW solution and weigh on a balance . Transect the pancreas from the junction of the head and tail of the pancreas, find the main pancreatic duct, insert a plastic tube, and perfuse a collagenase solution (1.5 g / L, containing DNase 0.3 g / L). Digest in a water bath at 37°C for 30 minutes. During the digestion process, the pH is about 7.5. From 20 minutes of digestion, samples are taken every 4 minutes. Dithizone staining is used for microscopic examination. When the pancreatic tissue is digested and cracked into fine sand, the microscopic examination shows that most of the structures are completed. The islets ...

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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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