Cell cultures of and cell culturing method for nontransformed pancreatic, thyroid, and parathyroid cells

A method for producing an expanded, enriched, non-transformed human cell culture of human cells selected from the group consisting of thyroid endocrine and parathyroid endocrine cells comprising:

• (a) selecting said cells from a human tissue that comprises said cells;
• (b) concentrating said cells selected in (a);
• (c) resuspending said cells concentrated in (b) in a growth medium;
• (d) culturing said cells resuspended in (c) in said growth medium to effect sustained cell division;
• and
• (e) passaging said cells cultured in (d) periodically to expand said culture;
• wherein said growth medium comprises a combination of a suitable basal medium and supplements of ingredients including about 0.48 mM MgCl.sub.2, about 0.01 mM hydrocortisone, about 2 ng/ml selenous acid, about 3 pg/ml triiodothyronine, about 75 .mu.g/ml bovine hypothalamus extract, about 5 .mu.g/ml bovine pituitary extract, and serum, which ingredients of the growth medium are present in concentrations effective to provide a medium which is selective for said human cells and in which said human cells are selectively proliferated without being transformed and differentiated functions are retained through periodic passaging, providing an expanded human cell culture of functionally differentiated, expanded, non-transformed human cells that is substantially free of passenger cells including fibroblast, macrophage, and capillary endothelial cells.

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

• (a) selecting said pancreatic endocrine cells from human pancreatic tissue;
• (b) concentrating said selected cells;
• (c) resuspending said concentrated cells in a growth medium;
• (d) culturing said resuspended cells to effect sustained cell division;
• and
• (e) passaging said cultured cells periodically to expand said culture;
• wherein said growth medium comprises a combination of a suitable basal medium and supplements of ingredients including hypothalamus and pituitary extracts, one or both of placental extract or human placental lactogen, and serum, which ingredients of said growth medium are adjusted in amounts to be effective to provide a medium which promotes sustained cell division of the cultured pancreatic endocrine cells and selects in favor of the cultured pancreatic endocrine cells and in which said cultured pancreatic endocrine cells are selectively proliferated without being transformed and differentiated functions are retained through periodic passaging, providing an expanded human pancreatic cell culture of functionally differentiated, expanded, non-transformed human pancreatic endocrine cells that is substantially free of passenger cells including fibroblast, macrophage, and capillary endothelial cells.

A method for producing an expanded, enriched, non-transformed human cell culture consisting essentially of human pancreatic endocrine cells comprising:

• (a) selecting human pancreatic endocrine cells from a human tissue;
• (b) concentrating said cells selected in (a);
• (c) resuspending said cells concentrated in (b) in a growth medium;
• (d) culturing said cells resuspended in (c) to effect sustained cell division;
• and
• (e) passaging said cells cultured in (d) periodically to expand said culture;
• wherein said growth medium comprises a basal medium and supplements of ingredients including hypothalamus and pituitary extracts, serum and calcium, which ingredients of said growth medium are adjusted in amounts to be effective provide a medium which is selective for said human cells and in which said human cells are selectively proliferated without being transformed and differentiated functions are retained through periodic passaging, providing an expanded human pancreatic endocrine cell culture that is substantially free of passenger cells including fibroblast, macrophage, and capillary endothelial cells.

A method for producing an expanded, enriched, non-transformed human cell culture of human thyroid endocrine or parathyroid endocrine cells comprising:

• (a) selecting said cells from a human tissue that comprises said cells;
• (b) concentrating said selected cells;
• (c) resuspending said concentrated cells in a growth medium;
• (d) culturing said resuspended cells in said growth medium to effect sustained cell division;
• and
• (e) passaging said cultured cells periodically to expand said culture;
• wherein said growth medium comprises a combination of a suitable basal medium and supplements of ingredients including MgCl.sub.2, hydrocortisone, selenous acid, triiodothyronine, hypothalamus extract, pituitary extract, and serum, which ingredients of the growth medium are adjusted in amounts to be effective to provide a medium which is selective for said human cells and in which said human cells are selectively proliferated without being transformed and differentiated functions are retained through periodic passaging, providing an expanded human cell culture of functionally differentiated, expanded, non-transformed human thyroid endocrine cells or parathyroid endocrine cells that is substantially free of passenger cells including fibroblast, macrophage, and capillary endothelial cells.