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Separation and extraction fluid suitable for pig pancreas islet and preparation and application thereof

A technology for extracting liquid and islets, which is applied in the field of separating and extracting liquid to improve the degree of hypoxia in separating and extracting islets, to achieve the effects of improving activity and function, increasing success rate, and avoiding damage

Inactive Publication Date: 2013-10-23
DALIAN INST OF CHEM PHYSICS CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although there are many studies on the isolation and extraction of islets, no researchers have developed an isolation and extraction solution that can effectively isolate islets and effectively improve the hypoxic state of islets.

Method used

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  • Separation and extraction fluid suitable for pig pancreas islet and preparation and application thereof

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Experimental program
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Effect test

Embodiment 1

[0038] Preparation and application of separation extract suitable for porcine islets:

[0039] RPMI1640 powder was stirred and dissolved with ultrapure water to prepare a concentrated solution so that the final concentration of the solute in the medium was twice the medium solution under standard conditions; the final concentration was 2 mg / m collagenase P (Roche Company, product number 11213873001) reagent was added , the final concentration is DNase 0.3mM, the final concentration is 5% bovine serum albumin, the final concentration is 200u / ml and 200ug / ml penicillin and streptomycin, after stirring evenly, 0.22um filter membrane is aseptically filtered, and the filtrate is taken.

[0040] Dissolve egg yolk lecithin with ultrapure water at a concentration of 5g / L, stir rapidly at room temperature, filter through a 0.22um sterile filter, and take the filtrate; perfluorotributylamine is directly filtered through a 0.22um filter membrane, and then take the filtrate and egg yolk L...

Embodiment 2

[0042] Preparation and application of separation extract suitable for porcine islets:

[0043] The Hank`s solution was stirred and dissolved with ultrapure water, and the final concentration of the solute in the prepared balanced salt solution was a salt solution that was 2 times under standard conditions; add 3 mg / ml collagenase P (Roche Company, product number 11213873001) reagent, DNase 0.6 mM, 4% bovine serum albumin, 200u / ml and 200ug / ml penicillin and streptomycin, stirred evenly and sterile filtered with 0.22um filter membrane. Dissolve soybean lecithin with ultrapure water at a concentration of 3g / L, stir rapidly at room temperature, and pass through 0.22um sterile filtration; perfluorotripropylamine is directly filtered through a 0.22um filter membrane and then mixed with soybean lecithin at a volume of 1:2 Ratio mixing, ultrasonic preparation of emulsion under sterile conditions, 200W, ultrasonic for 5 minutes. Mix the emulsion with balanced salt solution 1:1, stir ...

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Abstract

The invention provides a separation and extraction fluid suitable for pig pancreas islet and a preparation thereof. The fluid is characterized in that the oxygen load of the separation and extraction fluid is high and the problem of oxygen deficit in the separation and extraction process of pancreas islet can be solved. The separation and extraction fluid comprises the following steps of: first, taking a basal culture medium or a balanced salt solution as an aqueous phase; and adding collagenase, calcium chloride, DNA enzyme, bovine serum albumin, perfluorocarbon, an emulgator and an antibacterial agent for meeting the demand of separating and extracting pig pancreas islet. The separation and extraction fluid provided by the invention cannot release the anoxic degree in the separation and extraction process of pancreas islet and the survival rate, the function and the activity of the pancreas islet treated by the separation and extraction fluid provided by the invention are superior to those of pancreas islet treated by the conventional separation fluid with collagenase only.

Description

technical field [0001] The invention relates to a separation and extraction liquid product, specifically the separation and extraction liquid which can improve the degree of hypoxia in the separation and extraction of pancreatic islets. Background technique [0002] Pancreatic islet transplantation is a potential clinical treatment method for insulin-dependent diabetes. Compared with pancreas transplantation, islet transplantation has the advantages of simplicity, safety, effectiveness, and less side effects, so its application prospect is more extensive. However, the shortage of human-derived pancreas donors is the main problem restricting the treatment of islet transplantation, and pig islet transplantation is the best method to solve the source of islets [Moon-Kyu Lee. Cell transplantation for endocrine disorders, Advanced Drug Delivery Reviews, 2000, 42: 103-120] [Reinhard G. Islet cell transplantation today. Langenbecks Arch Surg, 2007, 392: 239-253]. However, when the...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/071
Inventor 马小军李娜于炜婷张英李珅张建斌马颖
Owner DALIAN INST OF CHEM PHYSICS CHINESE ACAD OF SCI
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