Methods of using genetically engineered cells that produce insulin in response to glucose
First Claim
1. A method of providing glucose-responsive insulin secreting capability to a mammal comprising implanting into a mammal in need of insulin secreting capability a therapeutically effective amount of a cell population positioned in a selectively permeable membrane, said cell population comprising engineered cells that secrete insulin in response to glucose, the engineered cells comprising a gene encoding low affinity, high Km glucokinase (hexokinase IV), an insulin gene and a GLUT-2 glucose transporter gene, wherein at least one of said genes is a recombinant gene introduced into the cell by means of a recombinant vector.
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Abstract
The present disclosure relates to the application of genetic engineering to provide artificial β cells, i.e. cells which can secrete insulin in response to glucose. This is achieved preferably through the introduction of one or more genes selected from the insulin gene, glucokinase gene, and glucose transporter gene, so as to provide an engineered cell having all three of these genes in a biologically functional and responsive configuration. Assays for detecting the presence of diabetes-associated antibodies in biological samples using these and other engineered cells expressing diabetes-associated epitopes are described. Also disclosed are methods for the large-scale production of insulin by perfusing artificial β cells, grown in liquid culture, with glucose-containing buffers.
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36 Claims
- 1. A method of providing glucose-responsive insulin secreting capability to a mammal comprising implanting into a mammal in need of insulin secreting capability a therapeutically effective amount of a cell population positioned in a selectively permeable membrane, said cell population comprising engineered cells that secrete insulin in response to glucose, the engineered cells comprising a gene encoding low affinity, high Km glucokinase (hexokinase IV), an insulin gene and a GLUT-2 glucose transporter gene, wherein at least one of said genes is a recombinant gene introduced into the cell by means of a recombinant vector.
Specification