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Last Updated: November 29, 2021

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Claims for Patent: 8,609,328

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Summary for Patent: 8,609,328
Title:Method of separating pancreatic islet
Abstract: The present invention provides a pancreatic islet isolation method comprising the steps of (1) injecting a protection solution containing a protease inhibitor into the pancreatic duct of an procured pancreas; (3) digesting the pancreas into which the protection solution has been injected; and (4) purifying the digested pancreatic tissue using a purification solution containing a density gradient reagent. The present invention also provides a protection solution for injection into the pancreatic duct, a pancreas preservation solution for the two-layer method, and an islet purification solution.
Inventor(s): Matsumoto; Shinichi (Kyoto, JP), Wada; Hiromi (Kyoto, JP), Noguchi; Hirofumi (Kyoto, JP)
Assignee: Otsuka Pharmaceutical Factory, Inc. (Naruto-shi, Tokushima, JP)
Application Number:13/624,309
Patent Claims:1. A method for isolating pancreatic islets comprising the sequential steps of: (a) injecting a protection solution containing a protease inhibitor and approximately 4 to 50 mmol/L of potassium into the pancreatic duct of a procured pancreas, wherein said protease inhibitor is at least one selected from the group consisting of ulinastatin, gabexate mesilate and nafamostat mesilate; (b) injecting a collagenase solution into said pancreas into which the protection solution has been injected and digesting the pancreas; (c) forming a centrifuged density gradient by centrifuging a density gradient obtained from a purification solution containing a density gradient reagent and to which tissue from said digested pancreas has been added, wherein said density gradient reagent is Ficoll solution or iodixanol; and (d) recovering and isolating said pancreatic islets from said centrifuged density gradient.

2. The isolation method according to claim 1, wherein in (a) the protection solution is injected into the pancreatic duct in an amount of approximately 0.1 to 10 ml per 1 gram organ weight.

3. The isolation method according to claim 1, wherein in (c) the purification solution further contains trehalose.

4. The isolation method according to claim 1, wherein in (c) the density gradient reagent is iodixanol.

5. The isolation method according to claim 1, wherein in (c) the purification solution further contains a protease inhibitor.

6. The method according to claim 1, wherein in (a) said protease inhibitor is ulinastatin.

7. The method according to claim 1, wherein in (a) said protease inhibitor is gabexate mesilate.

8. The method according to claim 1, wherein in (a) said protease inhibitor is nafamostat mesilate.

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