Neural cells expressing adenovirus E4ORF1, and methods of making and using the same
OwnerLiang Qiang, Daniel Joseph Nolan +1inventors
InventorsLiang Qiang, Daniel Joseph Nolan, Claude Geoffrey Davis
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Abstract
In certain aspects the present invention provides engineered neural cells, neural stem cells, or neural progenitor cells that contain a nucleotide sequence that encodes an adenovirus E40RF1 polypeptide and/or that contain an adenovirus E40RF1 polypeptide. The present invention also provides methods of making and using such engineered cells and compositions comprising such engineered cells.
Claims36
A substantially pure isolated population of engineered neural cells, engineered neural stem cells, or engineered neural progenitor cells, wherein the engineered neural cells, the engineered neural stem cells, or the engineered neural progenitor cells:
- a. comprise a nucleic acid sequence that encodes an adenovirus E40RF1 polypeptide wherein the nucleic acid is operably linked to a promoter,
- b. do not comprise an entire adenovirus E4 region, and
- c. do not comprise any E4ORF2, E4ORF3, E4ORF4, E4ORF5 or E4ORF6 coding sequences or amino acid sequences.
An in vitro blood brain barrier model system comprising endothelial cells and a population of engineered glial cells, wherein the engineered glial cells:
- (a) comprise a nucleic acid sequence that encodes an adenovirus E40RF1 polypeptide wherein the nucleic acid is operably linked to a promoter,
- (b) do not comprise an entire adenovirus E4 region, and
- (c) do not comprise any E4ORF2, E4ORF3, E4ORF4, E4ORF5 or E4ORF6 coding sequences or amino acid sequences.
An in vitro composition comprising neural cells and a carrier solution for delivery to a subject, wherein at least 75% of the neural cells in the composition are engineered neural cells that comprise a nucleic acid sequence that encodes an adenovirus E40RF1 polypeptide, do not comprise an entire adenovirus E4 region, and do not comprise any E40RF2, E40RF3, E40RF4, E40RF5 or E40RF6 coding sequences or amino acid sequences.
A method for culturing a population of neurons, the method comprising either:
- (a) culturing the population of neurons and a population of engineered glial cells together in the same culture vessel, wherein the engineered glial cells contain a nucleotide sequence that encodes an adenovirus E40RF1 polypeptide or contain an adenovirus E40RF1 polypeptide, or (b) contacting the population of neurons with glial-cell conditioned medium, wherein the glial-cell conditioned medium is obtained from a culture of the engineered glial cells thereby culturing the population of neurons.
Dependencies are inferred from the claim text.