Novel highly efficient method for preparing mice pancreatic single cell suspension

A single-cell suspension, pancreas technology, applied in animal cells, vertebrate cells, artificial cell constructs, etc., can solve the problems of difficult analysis, difficult separation and purification of pancreatic single cells, etc. The effect of high cell ratio

Inactive Publication Date: 2010-05-12
NORTHEAST FORESTRY UNIVERSITY +2
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Problems solved by technology

Compared with the traditional separation method, this invention has the ability to completely digest and separate the pancreas, and the obtained cell ratio is high, and the separated cells exist in a single form, have no adhesion, less impurities, and the separated

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  • Novel highly efficient method for preparing mice pancreatic single cell suspension

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Embodiment Construction

[0021] 1. Ultra-clean bench and sterile room are sterilized by ultraviolet radiation for 40 minutes, then turn off the ultraviolet equipment, open the exhaust system, and enter the sterile room after 5 minutes.

[0022] 2. Take 3 petri dishes with a diameter of 3.5 cm and place them on a clean bench. Pour about 2 mL of cold PBS (4°C) into two of the dishes, place them on ice packs, and set aside.

[0023] 3. Prepare two sets of clean surgical instruments, put them on the ultra-clean table, and set them aside.

[0024] 4. Kill a mouse by cervical dislocation, soak it in 75% ethanol for 3 minutes, put it into the sterile room after disinfection.

[0025] 5. Place the left abdomen of the mouse upwards and place it flat in a clean 10cm glass dish. Open the abdominal cavity of the mouse on the ultra-clean table to expose the internal organs. Carefully remove the pancreas and place it in a 3.5cm petri dish filled with cold PBS. middle.

[0026] 6. Move the small dish containing pa...

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Abstract

The invention discloses a novel highly efficient method for preparing a mice pancreatic single cell suspension. The method comprises the steps of selection of mouse pancreas, cold digestion of pancreatin, separation and purification of pancreatic single cells and the like. The key technology of the method is that: the pancreatin is used for invading the pancreatic tissue under conditions of low temperature and low activity, and the pancreatic tissue is dissociated by a short warm digestion so as to obtain the pancreatic single cell suspension with high quantity, high purification, high activity and no adhesion. Compared with the common pancreatic cell separation method, the method of the invention has the characteristics of stable operation, high cell yield, high cell activity and the like, avoids the defect existing in the common pancreatic cell separation method that the pancreatic cells are damaged due to long-time warm incubation of the pancreatin, the DNA released when the cells are died in large quantity is wound so as to cause the cell adhesion and the like, solves the problem of difficult separation and analysis of the pancreatic single cells and lays a foundation for further using the pancreatic cells.

Description

technical field [0001] The invention relates to a novel and efficient technology for separating and preparing mouse pancreatic single-cell suspension, in particular to a method for separating and preparing high-activity, high-purity, and non-adhesive pancreatic single-cell suspension by adopting a trypsin cold digestion method. Background technique [0002] Diabetes mellitus is a metabolic disease characterized by hyperglycemia caused by the lack or dysfunction of pancreatic β cells. In recent years, with the development of social economy and the improvement of residents' living standards in various countries in the world, the incidence and prevalence of diabetes have increased year by year. Diabetes not only affects the quality of life of patients and brings a heavy psychological burden to patients, but also the complications of diabetes pose a threat to the health and life of patients. [0003] Pancreatic stem cells are a type of pluripotent cells with high proliferation ...

Claims

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Application Information

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IPC IPC(8): C12N5/071
Inventor 滕春波李丹
Owner NORTHEAST FORESTRY UNIVERSITY
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