Method of isolating cells from a tissue in a mammal

Patent metadata

Field	Value
Publication number	US-9657271-B2
Application number	US-201313755091-A
Country	US
Kind code	B2
Filing date	Jan 31, 2013
Priority date	Jan 24, 2006
Publication date	May 23, 2017
Grant date	May 23, 2017

Abstract

Solutions and suspensions comprising polymerized hemoglobin derived from human blood are disclosed. The solutions and suspensions may comprise cell culture medium, an enzyme (such as a protease), and/or a buffer. Processes of preparing the solutions and suspensions are also disclosed. The solutions and suspensions may be employed in methods of isolating mammalian cells, such as pancreatic islets, methods of preserving mammalian tissue and organs, methods of aiding the recovery of mammalian cells following their isolation, methods of maintaining mammalian cells, methods of propagating mammalian cells, and methods of treating a mammal with diabetes.

First claim

What is claimed is:

1. A method of isolating pancreatic islets from a pancreatic tissue in a mammal, comprising the steps of:
a) contacting the pancreatic tissue of the mammal that includes the pancreatic islets a solution that includes polymerized hemoglobin derived from mammalian blood and a proteolytic enzyme; wherein the solution includes a concentration of between 5 and 15 g/dL of polymerized hemoglobin; and thereafter
b) recovering the pancreatic islets from the mammalian tissue, thereby isolating the pancreatic islets.

2. The method of claim 1 , wherein the tissue is human pancreatic tissue and the polymerized hemoglobin is derived from human blood.

3. The method of claim 2 , wherein the polymerized hemoglobin derived from human blood is pyridoxylated.

4. The method of claim 2 , wherein the solution is oxygenated.

5. The method of claim 2 , wherein the contacting step is achieved by intraductal administration.

6. The method of claim 2 , further including the step of washing the pancreatic islets with a cell culture medium to produce washed pancreatic islets.

7. The method of claim 6 , further including the step of purifying the washed pancreatic islets by density gradient centrifugation.

8. The method of claim 1 , wherein the solution further includes a cell culture medium.

9. The method of claim 8 , wherein the cell culture medium is RPMI.

10. The method of claim 1 , wherein the proteolytic enzyme is collagenase.

11. The method of claim 1 , wherein the solution further includes a buffer.

12. The method of claim 1 , wherein the polymerized hemoglobin is derived from human blood and is pyridoxylated.

13. The method of claim 12 , wherein the solution further includes a cell culture medium.

14. The method of claim 13 , wherein the proteolytic enzyme is collagenase and the cell culture medium is RPMI.

15. The method of claim 14 , wherein the solution is oxygenated.

16. The method of claim 1 , wherein the solution is oxygenated.

Assignees

• Hemoglobin Oxygen Therapeutics Llc
• Univ Illinois

Inventors

• Oberholzer Jose
• DOUBLEDAY Marc

Classifications

• C12N5/0676 Primary

  Pancreatic cells · CPC title

• A61K35/39

  Pancreas; Islets of Langerhans (Langerhans cells of epidermis A61K35/36) · CPC title

• A61K38/42

  Haemoglobins; Myoglobins · CPC title

• A61P3/10

  for hyperglycaemia, e.g. antidiabetics · CPC title

• A61K38/482

  Serine endopeptidases (3.4.21) · CPC title