Separation and purification method for islet cells of mouse

A technology for separation and purification of islet cells, which is applied in the field of cell separation, to achieve the effect of short digestion time of pancreas, simple and easy operation, and increased success rate of fractionation

Inactive Publication Date: 2013-04-03
SHANGHAI INST FOR ENDOCRINE & METABOLIC DISEASES
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Problems solved by technology

However, the CBD method cannot be used to isolate pancreatic islets in mice with abnormal pancreatic duct shape, young mice, high-fat-fed mice, pancreatitis and islet neonatal mouse models

Method used

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  • Separation and purification method for islet cells of mouse
  • Separation and purification method for islet cells of mouse
  • Separation and purification method for islet cells of mouse

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Embodiment 1

[0036] (1) Surgical operation: Mice were anesthetized by intraperitoneal injection of pentobarbital sodium solution (45 mg / kg body weight). Once fully anesthetized, 75% alcohol was used to disinfect the skin, and then an incision was made along the midline of the abdomen to fully expose the pancreas.

[0037] (2) Pancreas filling: cut off the diaphragmatic splenic ligament and gastrosplenic ligament to free the splenic pancreas, and cut the ligaments between the descending colon and rectum and pancreas at the same time; turn the splenic pancreas horizontally from left to right to clearly The superficial splenic vein is seen on the back of the pancreas. Clamp the junction of the splenic vein and spleen with a hemostat to prevent the collagenase solution from pouring into the spleen. The diaphragm was dissected, and the heart was cut open to terminate the blood supply to the pancreas; under a dissecting microscope, a 24G trocar was passed into the splenic vein at its junction wi...

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Abstract

The invention belongs to the technical field of cell separation, and particularly relates to a separation method for islet cells of a mouse. The method comprises the following steps: narcotizing the mouse, disinfecting the skin and cutting along the center of the abdomen to fully expose the pancreas; shearing the ligament organization connected with the pancreas of the spleen, clamping to close the junction of the splenic vein and the spleen and cutting the heart broken, inserting a needle at the intersection of the splenic vein and the portal vein to the splenic vein, and injecting collagenase filling liquid; immersing the filled pancreas of the spleen into the collagenase filling liquid after the pancreas of the spleen is dissociated from the abdominal cavity; performing water bath slaking on the pancreas collagenase solution, performing centrifugal separation after adding Hank's liquid and taking the sediment; and performing heavy-suspending sedimentation, filtration and centrifugal treatment on the sediment to obtain the islet cells of the mouse. The separation and purification method for the islet cells of the mouse, provided by the invention, is simple and feasible in operation, and the separated islet cells have no obvious differences with those obtained by a CBD method in forms and functions, so that the selectable technologies for separation of pancreas islet of the mouse are enriched, and the separative power is improved.

Description

technical field [0001] The invention belongs to the technical field of cell separation, in particular to a method for islet cell separation of mice. Background technique [0002] Diabetes has become the third biggest killer that threatens human health. The high blood sugar state of diabetic patients is conducive to the growth and reproduction of bacteria in the body, and it also inhibits the ability of white blood cells to phagocytose bacteria, reducing the patient's ability to resist infection. The common ones are urinary tract infection, respiratory tract infection, skin infection, nerve cells and nerve fibers are prone to lesions and so on. The chance of coronary heart disease in diabetic patients is 2 to 3 times that of non-diabetic patients. Retinopathy is also a common disease in diabetic patients. At present, among the more than 90 million diabetic patients in China, type 1 diabetes patients account for about 5%, and islet cell transplantation is one of the most effe...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/071
Inventor 宁光李峰曹亚南姜秀丽
Owner SHANGHAI INST FOR ENDOCRINE & METABOLIC DISEASES
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