Construction method of SD (Sprague Dawley) rat pancreas duct epithelial cell model

A technology of epithelial cells and construction methods, applied in the field of SD rat pancreatic ductal epithelial cell model construction, can solve the problems of low success rate, high equipment demand, low success rate, etc. The method is simple and easy, and the source is abundant and reliable , high success rate effect

Inactive Publication Date: 2017-03-22
覃蒙斌
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Problems solved by technology

The disadvantage of this method is that it is difficult to select the sieve, and the Ficoll solution needs to be freshly prepared, the equipment requirements for gradient centrifugation are high, the operation is cumbersome and the success rate is not high
The other is to directly isolate the pancreatic tissue of SD suckling mice, cut it into pieces, digest it with type IV collagenase, and collect the centrifuged sediment for p

Method used

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Embodiment Construction

[0022] Below by embodiment the present invention will be further described.

[0023] 1. Animal selection and anesthesia. One male SD rat with a body weight of 350 g was selected, and 10% chloral hydrate was used, and the ratio of the injection volume to the body weight of the SD rat was 0.3ml / 100g.

[0024] 2. Surgical separation of the main pancreatic duct in rats, the steps are (1) disinfect the abdomen three times with iodine and 75% alcohol; (2) open the abdominal cavity through a midline abdominal incision; (3) find the duodenum along the stomach, and turn out the pancreas ; (4) Identify and separate the bile duct and main pancreatic duct; (5) Cut off the main pancreatic duct, put it into a 30mm×10mm culture dish and add 2~3ml of PBS solution, and use iris scissors and micro forceps to separate and resect it with the naked eye The tissue surrounding the main pancreatic duct, the main pancreatic duct.

[0025] 3. Chemical digestion of the main pancreatic duct, put the mai...

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Abstract

The invention discloses a construction method of an SD (Sprague Dawley) rat pancreas duct epithelial cell model. The construction method adopts a rat and is characterized by further comprising steps as follows: animal selection and anesthesia, surgical separation of a main pancreatic duct of the rat, chemical digestion of the main pancreatic duct and cell type identification. According to the construction method, the main pancreatic duct is directly separated under the naked eyes according to a pancreatic anatomical structure of the SD rat, the separation recovery rate of epithelial cells of the pancreatic duct is higher, the obtained epithelial cells of the pancreatic duct are less irritated, and stable passage can be realized. The method is simple and convenient, the success rate is high, and the novel cell model with rich and reliable sources is provided for research of diseases of pancreas.

Description

technical field [0001] The invention relates to a cell model, especially a method for constructing a SD rat pancreatic ductal epithelial cell model. Background technique [0002] Pancreatic lesions are one of the diseases with the most complicated pathogenesis in human diseases. The inflammatory lesions are critical and difficult to treat; while the neoplastic diseases are difficult to detect early, with high mortality and poor treatment effect. Most pancreatic inflammation and tumors are caused by ductal epithelial cell lesions. However, it is very difficult to isolate and culture pancreatic ductal epithelial cells, which greatly limits the research on pancreatic lesions. At present, foreign studies on the in vitro culture of primary rat pancreatic ductal epithelial cells are mainly concentrated in the 1970s. The main method is: directly excise the head tissue of the rat pancreas, separate the main pancreatic duct area under a stereomicroscope, and place it in the After di...

Claims

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Application Information

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IPC IPC(8): C12N5/071
CPCC12N5/0676C12N2509/00
Inventor 覃蒙斌常仁杰梁志海唐国都
Owner 覃蒙斌
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