In vitro growth of functional islets of Langerhans and in vivo uses thereof
First Claim
1. A method for the in vitro growth of islet producing stem cells, IPSCs, to produce islet cells or an islet-like structure which comprises culturing pancreatic cells from a mammalian species in a basal nutrient medium supplemented with normal serum at below about 0.5% and glucose at below about 1 mM, allowing said IPSCs to grow for at least about 3 weeks, and initiating cellular differentiation into mature islet cells by re-feeding the IPSCs in culture with a nutrient medium supplemented with normal serum at about 0.5-10% and glucose at about 2.5 mM-10 mM.
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Abstract
The subject invention concerns new methods which make it possible, for the first time, to grow functional islets in in vitro cultures. The subject invention also concerns the use of the in vitro grown islet-like structures for implantation into a mammal for in vivo therapy of diabetes. The subject invention further concerns a process using the in vitro grown islet implants for growing an organ in vivo that has the same functional, morphological and histological characteristics as those observed in normal pancreatic tissue. The ability to grow these cells in vitro and organs in vivo opens up important new avenues for research and therapy relating to diabetes.
210 Citations
19 Claims
- 1. A method for the in vitro growth of islet producing stem cells, IPSCs, to produce islet cells or an islet-like structure which comprises culturing pancreatic cells from a mammalian species in a basal nutrient medium supplemented with normal serum at below about 0.5% and glucose at below about 1 mM, allowing said IPSCs to grow for at least about 3 weeks, and initiating cellular differentiation into mature islet cells by re-feeding the IPSCs in culture with a nutrient medium supplemented with normal serum at about 0.5-10% and glucose at about 2.5 mM-10 mM.
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18. A method for in vitro neogenesis of islet-like structures from pluripotent stem or progenitor cells which comprises:
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a. establishing a stromal, or "nurse", cell monolayer of ductal pancreatic epithelial cells which permits the generation of IPSCs; b. inducing stem/progenitor cell proliferation with culture conditions which promote cyclical regeneration of IPSCs and also prevent premature differentiation of the IPSC; and c. expanding and differentiating the IPSCs to produce an islet-like structure comprising α
, β
, and δ
cells. - View Dependent Claims (19)
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Specification