Method for producing self-reproducing mammalian pancreatic islet-like structures
First Claim
1. A method for producing mammalian pancreatic islet-like structures hereinafter called ILS'"'"'s, having histological characteristics and insulin-producing properties corresponding to those of natural isolated fetal pancreatic islets and islets obtained from adult animals maintained in culture, comprising establishing under conditions promoting the growth of mammalian cells, a culture comprising at least one pancreatic tissue-derived body attached to a substrate compatible with mammalian cells and capable of accepting the attachment of mammalian pancreatic islet tissue thereto, and immersed in a liquid tissue culture medium capable of sustaining mammalian cell growth, said body being selected from the group consisting of:
- (a) intact mammalian pancreatic islets isolated directly from postpartal pancreatic tissue and present in the culture at an islet population density of less than about 10 islets per ml of the liquid culture medium;
(b) pieces of mammalian postpartal pancreatic duct;
(c) clusters of coherent pancreatic cells obtained by mild digestion of pieces of whole mammalian postpartal pancreas with a solution of a connective tissue-lysing enzyme, said clusters being substantially free of pancreatic exocrine tissue;
(d) cells characterized by having an oblong flat configuration with highly serrated ends and tenacious attachment to the substrate and obtained by culturing a body selected from the group consisting of said bodies (a), (b), and (c) above and (e) below for prolonged periods in said culture under said mammalian cell-growth promoting conditions;
(e) ILS'"'"'s obtained by culturing a body selected from the group consisting of said bodies (a), (b), (c), and (d) for prolonged periods under said mammalian cell-growth promoting conditions; and
maintaining said culture for a period sufficient to permit neogenesis of discrete ILS'"'"'s connected to cell tissue outgrowing from the body and attached to the substrate;
detaching said ILS'"'"'s from the said outgrowing tissue; and
recovering said detached ILS'"'"'s from the culture.
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Abstract
Method for producing pancreatic islet-like structures (ILS'"'"'s) having histology and insulin-producing properties corresponding to those of fetal pancreatic islets and islets from adult animals maintained in culture. The ILS'"'"'s are produced by culturing isolated natural pancreatic islets, pancreatic duct pieces, cell clusters consisting of mildly digested pieces of pancreas, cell tissues obtained as by-products of the culturing methods, or previously-produced ILS'"'"'s, in attachment to a substrate.
77 Citations
50 Claims
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1. A method for producing mammalian pancreatic islet-like structures hereinafter called ILS'"'"'s, having histological characteristics and insulin-producing properties corresponding to those of natural isolated fetal pancreatic islets and islets obtained from adult animals maintained in culture, comprising establishing under conditions promoting the growth of mammalian cells, a culture comprising at least one pancreatic tissue-derived body attached to a substrate compatible with mammalian cells and capable of accepting the attachment of mammalian pancreatic islet tissue thereto, and immersed in a liquid tissue culture medium capable of sustaining mammalian cell growth, said body being selected from the group consisting of:
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(a) intact mammalian pancreatic islets isolated directly from postpartal pancreatic tissue and present in the culture at an islet population density of less than about 10 islets per ml of the liquid culture medium; (b) pieces of mammalian postpartal pancreatic duct; (c) clusters of coherent pancreatic cells obtained by mild digestion of pieces of whole mammalian postpartal pancreas with a solution of a connective tissue-lysing enzyme, said clusters being substantially free of pancreatic exocrine tissue; (d) cells characterized by having an oblong flat configuration with highly serrated ends and tenacious attachment to the substrate and obtained by culturing a body selected from the group consisting of said bodies (a), (b), and (c) above and (e) below for prolonged periods in said culture under said mammalian cell-growth promoting conditions; (e) ILS'"'"'s obtained by culturing a body selected from the group consisting of said bodies (a), (b), (c), and (d) for prolonged periods under said mammalian cell-growth promoting conditions; and
maintaining said culture for a period sufficient to permit neogenesis of discrete ILS'"'"'s connected to cell tissue outgrowing from the body and attached to the substrate;
detaching said ILS'"'"'s from the said outgrowing tissue; and
recovering said detached ILS'"'"'s from the culture. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47)
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48. Method as claimed in 47 wherein the medium contains about 100 μ
- U/ml penicillin and about 100 μ
g/ml streptomycin.
- U/ml penicillin and about 100 μ
- 49. A method for producing mammalian pancreatic islet-like structures hereinafter called ILS'"'"'s, having histological characteristics and insulin-producing properties similar to those of natural isolated fetal pancreatic islets and islets obtained from adult animals maintained in culture, comprising sub-culturing neogenetic ILS'"'"'s harvested from a culture of pancreatic tissue-derived bodies, and harvesting neogenetic ILS'"'"'s from the sub-culture.
Specification