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Producing method of repairable pigling pancreatic island cell

A technology of islet cells and production method, applied in the field of biological cells, can solve problems such as poor culture effect and complex serum composition, and achieve the effects of easy mutagenesis, low immune antigenicity and high safety

Active Publication Date: 2009-10-28
张俊英
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the composition of serum is complex, and it is easy to cause poor culture effect

Method used

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  • Producing method of repairable pigling pancreatic island cell
  • Producing method of repairable pigling pancreatic island cell

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Experimental program
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Effect test

Embodiment Construction

[0039] specific implementation plan

[0040] The present invention is further described as follows in conjunction with specific step process below:

[0041] 1. Selection of heterogeneous islets:

[0042] 1. Characteristics of newborn piglet islets:

[0043](1) Both human and porcine insulin molecules have chain A and chain B, both of which are composed of 51 amino acids. The A chain contains 21 amino acids, and the B chain contains 30 amino acids. And there are disulfide chain connections between A and B chains. The structure is very similar, the difference is only one amino acid difference. The 30th amino acid in the B chain of human insulin is threonine. The 30th amino acid in the B chain of porcine insulin is alanine. (2) Porcine insulin has been used in the clinical treatment of patients with type 1 diabetes for more than ten years, which has fully proved the safety and effectiveness of porcine insulin in humans. (3) The immunogenicity of newborn pig islets is weak,...

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Abstract

A production method capable of repairing piglet islet cells, characterized in that it includes the following process steps: (1) donor selection: the donor selects healthy piglets that have just been born and have not been breastfeeding, and the pancreas is removed by aseptic laparotomy , blood vessels, and connective tissue were placed in Hank`s solution at 4°C; (2) islet cell isolation and purification: the islets were separated from exocrine tissue by mechanical grinding. Then through Dextran discontinuous density gradient centrifugation to obtain islet cells; (3) Islet cell culture: use serum-free microsphere carrier for CO2 culture, use RPMI-1640 culture medium, and add fibronectin. The cells are wrapped in APA biofilm to form microspheres, cultured in a CO2 incubator at a constant temperature of 37°C, and the culture medium is changed every 48 hours, and finally microencapsulated newborn piglet islet cells with good activity and high purity are obtained.

Description

technical field [0001] The invention relates to the technical field of biological cells, in particular to a production method of repairable piglet islet cells, which can be clinically applied to the islet transplantation of diabetic patients. Background technique [0002] Diabetes is a global disease that endangers human health. The morbidity rate is 3-5%; the mortality rate is second only to cardiovascular diseases and tumors. It is predicted that by 2025, there will be 300 million people with diabetes in the world. Among them, patients with type I diabetes (IDDM) will account for 5-10%; these people will depend on exogenous insulin for life. Long-term injection of insulin can cause local skin redness, swelling, weeping, induration, infection, subcutaneous fat atrophy or fibrotic hyperplasia. Daily insulin injections not only bring inconvenience and pain to patients, but more importantly, the application of insulin can only control hyperglycemia and cannot prevent the oc...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/06C12N5/071
CPCC12N5/0676
Inventor 张俊英王斌王章存范清堂
Owner 张俊英