Method of stimulating proliferation and differentiation of human fetal pancreatic cells ex vivo

• US 5,587,309 A
• Filed: 04/29/1994
• Issued: 12/24/1996
• Est. Priority Date: 04/29/1994
• Status: Expired due to Term

• Current Assignee: The Whittier Institute For Diabetes And Endocrinology, United States Department Of Health And Human Services
• Original Assignee: The Whittier Institute For Diabetes And Endocrinology, United States Department Of Health And Human Services
• Inventors: Otonkoski, Timo P. J., Hayek, Alberto, Rubin, Jeffrey, Beattie, Gillian M.
• Primary Examiner(s): Wityshyn, Michael G.
• Assistant Examiner(s): DADIO, SUSAN
• Application Number:
  US08/235,394
• Time in Patent Office: 970 Days
• Field of Search: 435/240.2, 435/240.21, 435/240.3, 435/240.31, 435/240.25
• US Class Current: 435/375
• CPC Class Codes: A61K 35/12 Materials from mammals; Compositions comprising non-specified tissues or cells; Compositions comprising non-embryonic stem cells; Genetically modified cells vaccines or medicinal preparations containing antigens or antibodies A61K39/00
• CPC Class Codes: A61P 3/08 for glucose homeostasis pancreatic hormones A61P5/48
• CPC Class Codes: C12N 2501/12 Hepatocyte growth factor [HGF]
• CPC Class Codes: C12N 5/0676 Pancreatic cells

Abstract

A method of inducing the proliferation and/or differentiation of human fetal pancreatic cells entails contacting such cells in primary culture with Hepatocyte Growth Factor/Scatter Factor, thereby inducing a proliferation of β-epithelial cells, an increase in the number of β-epithelial cells which form islet-like cell clusters, and an increase in insulin production per cell. The method provides increased numbers of functional islet-like cell clusters for transplantation, for example, into Type 1 diabetic patients. The method can be scaled up so as to provide clinically useful numbers of cells for transplantation.

Claims

1. A method for inducing the ex vivo proliferation and differentiation of human fetal pancreatic β

-islet cells, comprising the steps of;

(a) preparing a primary culture of human fetal pancreatic cells; and

(b) contacting said primary culture with an effective concentration of Hepatocyte Growth Factor/Scatter Factor, wherein said effective concentration is an amount sufficient to induce said primary culture to proliferate, to form insulin-producing islet-like cell clusters that upon grafting into a host animal develop into functional islet tissue, and to increase the proportion of β

-epithelial cells in said clusters.

2. The method of claim 1, wherein said cell proliferation and differentiation comprise an increase in insulin production per cell.

3. A method of claim 1, wherein said effective concentration of Hepatocyte Growth Factor/Scatter Factor ranges from about 5 to about 50 ng/ml.

4. The method of claim 1, further comprising contacting said primary culture with an amount of anti-Tumor Growth Factor β

antibody sufficient to increase cell proliferation and formation of islet-like pancreatic cell clusters.

5. The method of producing proliferating and differentiating human fetal pancreatic islet cells in clinically useful quantities, comprising the steps of:

(a) preparing a primary culture of human fetal pancreatic cells;

(b) seeding said primary culture into a bioreactor;

(c) perfusing said bioreactor with a complete growth medium supplemented with an effective concentration of Hepatocyte Growth Factor/Scatter Factor, wherein said effective concentration is an amount sufficient to induce said primary culture of human fetal pancreatic cells to proliferate, to form insulin-producing islet-like cell clusters that upon grafting into a host animal produces functional islet tissue, and to increase the proportion of β

-epithelial cells in said clusters; and

(d) harvesting said islet-like cell clusters.

6. The method of claim 5, further comprising contacting said primary culture with an amount of anti-Tumor Growth Factor β

antibody sufficient to increase cell proliferation and formation of islet-like pancreatic cell clusters.