US 5,646,035 A · Method for preparing an expanded culture and clonal strains of pancreatic, thyroid or parathyroid cells

Abstract

The present invention provides a method for producing an expanded non-transformed cell culture comprising the steps of: (1) preparing partially purified, minced tissue; (2) concentrating the resulting cells and tissue pieces; (3) resuspending the concentrated tissue cells and pieces in a culture medium capable of supporting sustained cell division that is contained in a culture vessel; (4) incubating the cells; and (5) passaging the cells periodically. The present invention further provides clonal strains of cells derived from the above-mentioned cell culture, medium and conditioned medium designed for the culturing of such cells, including pancreatic, thyroid, parathyroid, and parotid cells, and the use of cultured pancreatic cells to form pancreatic pseudotissues composed of matrix-embedded aggregated (pseudoislets) or individual cells, to treat blood sugar disorders in mammals, and to test for cytotoxicity and autoimmune activities with reference to pancreatic endocrine cells.

Claims16

Dependent claims

A method for producing an expanded, enriched, non-transformed cell culture of mammalian pancreatic endocrine cells, comprising the steps of:

(a) selecting said pancreatic endocrine cells from pancreatic tissue;
(b) concentrating said selected cells;
(c) resuspending said concentrated cells in a culture medium;
(d) culturing said resuspended cells for a time and under conditions to effect sustained cell division;
and
(e) passaging said cultured cells periodically to expand said culture;
wherein said culture medium is Coon's 4506.035 or Coon's 4506.07, and said culture is substantially free of fibroblast, macrophage, and capillary endothelial cells.

4 more elements

A method for producing an expanded, non-transformed cell culture of mammalian thyroid or parathyroid cells, comprising the steps of:

(a) selecting said thyroid or parathyroid cells from thyroid or parathyroid tissue;
(b) concentrating said selected cells;
(c) resuspending said concentrated cells in a culture medium;
(d) culturing said resuspended cells for a time and under conditions to effect sustained cell division;
and
(e) passaging said cultured cells periodically to expand culture;
wherein said culture medium is Coon's 4506.035 or Coon's 4506.07, modified so that the concentration of MgCl.sub.2 is about 0.48 mM, the concentration of hydrocortisone is 3.5 ng/ml, the concentration of selenous acid is about 2 ng/ml, the triiodothyronine concentration is about 3 pg/ml, the final concentration of bovine hypothalamus extract is about 75 .mu.g protein/ml, and the final concentration of bovine pituitary extract is about 5 .mu.g protein/ml, and said culture is substantially free of fibroblast, macrophage, and capillary endothelial cells.

4 more elements

Dependencies are inferred from the claim text.

Method for preparing an expanded culture and clonal strains of pancreatic, thyroid or parathyroid cells

Life of this patent

Method for preparing an expanded culture and clonal strains of pancreatic, thyroid or parathyroid cells

Life of this patent

Abstract

Claims16