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Method for expanding pancreatic beta cell in vitro

An in vitro expansion and β-cell technology, applied in the field of cell culture, can solve the problems of slow proliferation and small number of cells, and achieve the effects of simplifying operations, saving costs, and saving separation and purification steps

Active Publication Date: 2013-09-18
吉林省合众生物科技有限公司
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  • Abstract
  • Description
  • Claims
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AI Technical Summary

Problems solved by technology

Stem cell-induced differentiation of islet β cells also has the disadvantages of small number of cells and slow proliferation. Therefore, finding drugs that can promote the proliferation of islet β cells in vivo or in vitro is of great significance for the treatment of diabetes

Method used

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  • Method for expanding pancreatic beta cell in vitro
  • Method for expanding pancreatic beta cell in vitro
  • Method for expanding pancreatic beta cell in vitro

Examples

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Embodiment 1

[0055] 1. Isolation and purification of rat pancreatic ductal epithelial cells

[0056] A 7-week-old, 220g male wistar rat (Zhejiang Academy of Medical Sciences Animal Center) was fasted overnight before the operation, anesthetized by intraperitoneal injection of 200 μl of 1 mg / ml pentobarbital sodium, and the abdominal cavity was opened layer by layer through a median abdominal incision. The common bile duct was ligated at the portal end, and 3 mL of 1.5 mg / mL type IV collagenase (sigma) was injected antegradely from the end close to the duodenum, and the entire dilated pancreas was quickly removed. Wash with cold D-Hanks solution, cut the pancreatic tissue into small pieces, put it into a 15ml centrifuge tube, add 5ml of 1.5mg / mL collagenase V (sigma), and place it in a 37°C water bath for 15min for digestion. Add cold Hanks solution containing 10% calf serum to the centrifuge tube to stop the digestion, filter through an 80-mesh stainless steel screen, collect the filtrate,...

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Abstract

The invention discloses a method for expanding pancreatic beta cells in vitro. The method comprises the following steps of: (1) preparing expression supernatant of Reg Ialpha protein; (2) separating and purifying pancreas conduit epithelial cells, and carrying out induced differentiation to obtain pancreatic beta cells; and (3) adding the expression supernatant of the Reg Ialpha protein prepared in the step (1) into a cell culture medium, and inoculating the pancreatic beta cells to culture. The method for expanding pancreatic beta cells in vitro can be used for establishing an expansion model of the in-vitro expanded pancreatic beta cells, is helpful for screening pancreatic beta cell expansion prompting medicines in large scale and obtaining medicines for expanding the pancreatic beta cells in vivo and in vitro effectively, in particular Chinese patent medicines, so that pancreatic beta cells are enriched through in-vitro expansion to provide donorcells for diabetes treatment, or the pancreatic beta cell expansion condition is improved under the conditioning of the Chinese patent medicines.

Description

technical field [0001] The invention belongs to the technical field of cell culture, and in particular relates to a method for expanding pancreatic beta cells in vitro. Background technique [0002] Diabetes is a metabolic disease with multiple etiologies. Its prevalence has risen sharply in recent years, and it has become the third major killer of human health after tumors and cardiovascular diseases. [0003] The treatment of diabetes is still a life-long treatment. Type Ⅰ and some type Ⅱ diabetes patients rely on regular injection of insulin every day to maintain blood sugar stability. Long-term injection has brought great pain and trouble to patients. Now the methods of islet transplantation and cell therapy have brought hope for the treatment of diabetes. However, the limited source of human islet tissue inhibits the development and application of these treatments. For example, there are about 20,000 new cases of type 1 diabetes in the United States every year, but th...

Claims

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Application Information

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IPC IPC(8): C12N5/071
Inventor 贾静王孝举
Owner 吉林省合众生物科技有限公司
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