Method for constructing tissue engineering islet by adipose derived mesenchymal stem cells and obtained tissue engineering islet

A technology of adipose stem cells and mesenchymal stem cells, which is applied in the field of tissue engineering islets, can solve the problems of little research on islet tissue raw materials, reduce the success rate of tissue construction, and loss of seed cells, so as to improve cell utilization, promote cell adhesion, increase The effect of adsorption quantity

Inactive Publication Date: 2015-07-22
UNION STEMCELL & GENE ENG
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] Acellular matrix is ​​a natural biological material. The original acellular dermal matrix was used as a dermal substitute in burn surgery, neurosurgery and other fields. With continuous development, acellular matrix has developed autologous and allogeneic and a variety of species from different sources, but there are few studies on the use of islet tissue as a raw material for the preparation of acellular matrix
At the same time, in the process of using acellular matrix to construct engineered tissues, there is a defect that the seed cells are lost in the process of replacing the seed cells with the acellular matrix in the liquid medium, which reduces the success rate of tissue construction.

Method used

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  • Method for constructing tissue engineering islet by adipose derived mesenchymal stem cells and obtained tissue engineering islet
  • Method for constructing tissue engineering islet by adipose derived mesenchymal stem cells and obtained tissue engineering islet
  • Method for constructing tissue engineering islet by adipose derived mesenchymal stem cells and obtained tissue engineering islet

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Experimental program
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Embodiment 1

[0030] 1. In vitro isolation, culture and expansion of adipose-derived mesenchymal stem cells

[0031] 1) Weigh 7.6g of human adipose tissue obtained from surgery (discarded), wash it with phosphate buffered solution (PBS) containing 1000u / ml penicillin / streptomycin until clarified, and then cut the adipose tissue into chyle shape, Add an equal volume of PBS containing 3000u / ml type I collagenase, 37°C, 100 rpm, shaking and digesting for 45min;

[0032] 2) Wash twice with PBS after digestion, discard the supernatant after centrifugation for 5 minutes, resuspend the cells with adipose-derived stem cell culture medium, add them to T-75 cell culture flasks and culture them in a cell culture incubator, observe under the microscope after 24 hours A large number of cells adhere to the wall and grow, aspirate and discard the medium, wash once with 10ml PBS and add 15ml fresh medium to continue culturing;

[0033] 3) After 72 hours of inoculation, the cells grow to about 90%, wash th...

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Abstract

The invention discloses a method for constructing tissue engineering islet by adipose derived mesenchymal stem cells and the obtained tissue engineering islet. The method employs adipose derived mesenchymal stem cells with extensive sources to perform culture on islet acellular matrix, so that the acellular matrix can provide a three-dimensional space suitable for growth to adipose derived mesenchymal stem cells so as to promote cell adhesion, proliferation and differentiation, and formation of a three-dimensional structure engineering islet tissue able to stably secrete insulin. The method realizes gathering of insulin secreting cells, enhances cell utilization and is expected to promote the effect of repairing islet injury by tissue engineering method.

Description

technical field [0001] The invention relates to tissue-engineered islets, in particular to a method for constructing tissue-engineered islets using adipose-derived mesenchymal stem cells and the obtained tissue-engineered islets. Background technique [0002] Type I diabetes, also known as insulin-dependent diabetes mellitus, is an autoimmune disease characterized by selective destruction of islet β cells mediated by T lymphocytes, insufficient insulin secretion, and unbalanced regulation of blood sugar concentration, leading to hyperglycemia. At present, oral hypoglycemic drugs and exogenous insulin injections are the main methods for the treatment of type Ⅰ diabetes, but they cannot fundamentally cure diabetes. Islet transplantation is an effective way to completely cure diabetes, but it has been facing two key problems of insufficient donor sources and immune rejection. [0003] Adipose-derived mesenchymal stem cells are cells with multi-directional differentiation poten...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/071C12N5/0775A61L27/38A61L27/36A61L27/50
Inventor 陈晓波韩俊领韩洪起洪敬欣靳霞胡艳华张宇光王雪莲
Owner UNION STEMCELL & GENE ENG
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