PHOSPHATIDYLINOSITOL 3-KINASE INHIBITORS AS STIMULATORS OF ENDOCRINE DIFFERENTIATION
First Claim
1. A method of inducing differentiation of endocrine cells, said method comprising culturing a mammalian endocrine precursor cell in the presence of a phosphatidylinositol 3-kinase (PI3K) inhibitor whereby said endocrine precursor cell differentiates into a cell having endocrine activity.
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Abstract
This invention pertains to the discovery that inhibition of phosphatidylinositol 3-kinase (PI3K) in human fetal undifferentiated cells induces morphological and functional endocrine differentiation. This is associated with an increase in MRNA levels of insulin, glucagon, and somatostatin, as well as an increase in the insulin protein content and secretion response to secretagogues. Blockade of PI3K also increases the proportion of pluripotent precursor cells coexpressing multiple hormones and the total number of terminally differentiated cells originating from these precursor cells. In one embodiment, this invention thus rpov9des methods for endocrine differentiation in cultured cells.
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Citations
41 Claims
- 1. A method of inducing differentiation of endocrine cells, said method comprising culturing a mammalian endocrine precursor cell in the presence of a phosphatidylinositol 3-kinase (PI3K) inhibitor whereby said endocrine precursor cell differentiates into a cell having endocrine activity.
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8. A nutrient medium for use in the culture of differentiated mammalian cells having endocrine activity, said medium comprising:
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a mammalian cell culture medium; and
a phosphatidylinositol 3-kinase inhibitor. - View Dependent Claims (9, 10, 11, 12, 13, 14, 16, 17, 18, 19, 20, 21, 22, 23, 24, 26, 27, 28, 29, 30, 31, 32, 33, 34, 36, 37, 38, 39, 40)
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15. A process for obtaining animal cells by cell culture, said process comprising:
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(i) culturing mammalian precursor cells in a nutrient medium contained in a culture vessel, wherein said nutrient medium comprises a a mammalian cell culture medium; and
a phosphatidylinositol 3-kinase inhibitor;
(ii) continuing the culture in said nutrient medium until said precursor cells differentiate and have endocrine activity; and
(iii) collecting the differentiated cells.
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25. A bioreactor comprising a container containing
a nutrient medium, said nutrient medium comprising a mammalian cell culture medium; - and
a phosphatidylinositol 3-kinase inhibitor; and
a mammalian precursor cell capable of endocrine activity when differentiated.
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35. A method of treating a hormone deficiency in an organism, said method comprising
culturing a mammalian precursor cell in the presence of a phosphatidylinositol 3-kinase (PI3K) inhibitor whereby said precursor cell differentiates into a cell having endocrine activity; - and
transplanting said cell having endocrine activity into said organism.
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41. A kit for the in vitro culture of a differentiated endocrine cell, said kit comprising a container containing a phosphatidylinositol 3-kinase (PI3K) inhibitor and one or more other components selected from the group consisting of cell culture media, adult mammalian cells, fetal mammalian cells, instructional materials teaching the use of PI3K inhibitors to enhance the differentiation of endocrine cells in culture.
Specification