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Culturing pancreatic stem cells having a specified, intermediate stage of development

  • US 20020081725A1
  • Filed: 06/29/2001
  • Published: 06/27/2002
  • Est. Priority Date: 06/30/2000
  • Status: Active Grant
First Claim
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1. A method of preparing a cell culture of propagating pancreatic cells having the ability to be passed from one culture vessel to a second vessel at an initial concentration of about 180 cells per square centimeter and expanded to about 1,800 cells per square centimeter, while retaining the following properties:

  • 90% PDX-1 positive and having an insulin;

    actin mRNA ratio of between 1;

    100 and 1000;

    1, said method comprising the steps of;

    (a) isolating propagating pancreatic cells;

    (b) transferring the cells to a culture medium containing growth hormone and having 1% or less total volume of serum to selectively propagate cells having an insulin;

    actin mRNA ratio of between 1;

    100 and 1000;

    1 and that are PDX-1 positive; and

    (c) passaging the cells to provide a cell culture of propagating pancreatic cells having the ability to be passed from one culture vessel to a second vessel at an initial concentration of about 180 cells per square centimeter and expanded to about 1,800 cells per square centimeter while retaining the following properties;

    90% PDX-1 positive and having an insulin;

    actin mRNA ratio of between 1;

    100 and 1000;

    1.

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