Large scale pancreatic islet purification
AssigneeBaylor Research Institute
InventorsShinichi Matsumoto, Masayuki Shimoda
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Abstract
The present invention includes a method of isolating pancreatic islets by density centrifugation wherein the pancreatic islets are loaded in a solution comprising pancreatic islets with the density gradient and the islets are isolated by centrifuging the vessel wherein the pressure on the pancreatic islets is less than 50 Pa and wherein the pancreatic islets are isolated from the gradient, the improvement comprising creating a continuous density gradient in a vessel comprising at least 100 milliliters with a bend tube that reduces convection currents that disrupt the gradient.
Claims32
A method of isolating pancreatic islets comprising:
- creating a continuous density gradient in a vessel comprising at least 100 milliliters with a bend tube that reduces convection currents that disrupt the gradient;
- loading a solution comprising pancreatic islets in the density gradient;
- centrifuging the vessel wherein the pressure on the pancreatic islets is less than 50 Pa;
- and
- isolating the pancreatic islets from the gradient.
A method of isolating pancreatic islets comprising:
- creating a continuous density gradient in a vessel comprising at least 100 milliliters with a bend tube that reduces convection currents that disrupt the gradient, the continuous density solution, further defined as comprising a low-density (density:
- 1.077) to a high-density (density:
- 1.095-1.125) prepared using a bend tube;
- loading pancreatic islet cells in a dilution solution on the gradient;
- centrifuging the vessel wherein the pressure on the pancreatic islets is less than 50 Pa;
- separating into two or more collection vessels the gradient from the top to the bottom of the vessel;
- selecting the collection vessel with the pancreatic islets;
- and
- isolating the pancreatic islets from the gradient.
A method of isolating pancreatic islets comprising:
- creating a density gradient in a vessel comprising at least 100 milliliters;
- loading a solution comprising pancreatic islets in a dilution solution below the density gradient;
- centrifuging the vessel wherein the pressure on the pancreatic islets is less than 50 Pa;
- separating into two or more collection vessels the gradient from the top to the bottom of the vessel;
- selecting the collection vessel with the pancreatic islets;
- and
- isolating the pancreatic islets from the gradient.
A method of isolating pancreatic islets by density centrifugation wherein the pancreatic islets are loaded in a solution comprising pancreatic islets with the density gradient and the islets are isolated by centrifuging the vessel wherein the pressure on the pancreatic islets is less than 50 Pa and wherein the pancreatic islets are isolated from the gradient, the improvement comprising creating a continuous density gradient in a vessel comprising at least 100 milliliters with a bend tube that reduces convection currents that disrupt the gradient.
One or more pancreatic cells isolated by the method comprising:
- creating a continuous density gradient in a vessel comprising at least 100 milliliters with a bend tube that reduces convection currents that disrupt the gradient;
- loading a solution comprising pancreatic islets in the density gradient;
- centrifuging the vessel wherein the pressure on the pancreatic islets is less than 50 Pa;
- and
- isolating the pancreatic islets from the gradient.
Dependencies are inferred from the claim text.