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Neural cells expressing adenovirus E4ORF1, and methods of making and using the same

A technology of nerve cells and neural stem cells, applied in the field of nerve cells expressing adenovirus E4ORF1 and its preparation and application

Active Publication Date: 2017-04-19
ANGIOCRINE BIOSCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, to the applicant's knowledge, prior to the present invention, there was no evidence that the adenoviral E4 region, and especially the E4ORF1 sequence, might have any beneficial effects on neuronal or glial cells

Method used

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  • Neural cells expressing adenovirus E4ORF1, and methods of making and using the same
  • Neural cells expressing adenovirus E4ORF1, and methods of making and using the same
  • Neural cells expressing adenovirus E4ORF1, and methods of making and using the same

Examples

Experimental program
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Embodiment 1

[0071] Example 1: Passaging and Characterization of E4ORF1 Expressing Astrocytes

[0072] Methods: 10-12 neonatal mice (P1) were sacrificed and their brains were isolated into dissection medium. Separate the two cerebral hemispheres with forceps and carefully remove the meninges and olfactory bulbs. Mince the tissue into small pieces (approximately 0.5 mm in diameter) using fine surgical scissors. Tissue fragments were collected and transferred to a 15 ml conical tube along with dissection medium. After the tissue fragments have sat for approximately 1-2 minutes, remove them from the dissection medium. Add 6-8ml TrypLE to the conical tube and cap tightly. After inverting the conical tube up and down three times, place it in a 37°C water bath for 15 minutes, during which time the conical tube was vortexed intermittently. Without disturbing the tissue fragments, remove TrypLE and then add 1-2 ml of astrocyte medium. The addition of deoxyribonuclease (DNAse) was continued ...

Embodiment 2

[0075] Example 2: Cell generation and characteristics of neuronal cells expressing E4ORF1

[0076] METHODS: Day 18 mouse embryos (E18) were sacrificed and their brains were isolated into dissection medium. Separate the two cerebral hemispheres with forceps and carefully remove the meninges and olfactory bulbs. Carefully separate the hippocampus from the brain hemispheres. Mince the hippocampal tissue into small pieces (approximately 0.5 mm in diameter) using fine surgical scissors. Tissue fragments were collected and transferred to a 15 ml conical tube along with dissection medium. After the tissue fragments have sat for approximately 1-2 minutes, remove them from the dissection medium. Add 6-8ml TrypLE to the conical tube and cap tightly. After inverting the conical tube up and down three times, place it in a 37°C water bath for 15 minutes, during which time the conical tube was vortexed intermittently. Without disturbing the tissue fragments, remove TrypLE and then ad...

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Abstract

The present invention discloses 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 the engineered cells and compositions comprising the engineered cells.

Description

[0001] Cross References to Related Applications [0002] This application claims priority to US Provisional Patent Application No. 62 / 018,056, filed June 27, 2014, the contents of which are incorporated herein by reference. [0003] incorporated by reference [0004] To the extent only jurisdictions permit incorporation by reference, all documents cited herein are incorporated by reference in their entirety. Background technique [0005] Adenovirus early transcribed region 4 (E4) contains at least 6 open reading frames (E4 ORFs). Past studies have shown that the entire E4 region regulates angiogenesis and promotes endothelial cell survival (see Zhang et al. (2004), J. Biol. Chem. 279(12): 11760-66). Past studies have also shown that throughout the E4 region, the E4ORF1 sequence is responsible for these biological effects in endothelial cells. See US Patent No. 8,465,732, and Seandel et al. (2008), "Generation of functional and durable vascular niche by the adenoviral E4ORF1...

Claims

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Application Information

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IPC IPC(8): C12N5/0735
CPCA61K48/005C12N5/0619C12N2710/10333C12N2501/998C12N2510/00C12N5/0622A61P25/00A61P25/28A61K35/30C07K14/005C07K14/00C12N7/00C12N15/86C12N2502/081C12N2502/086C12N2506/00C12N2533/90
Inventor 强梁克劳德·杰弗里·大卫丹尼尔·约瑟夫·诺兰
Owner ANGIOCRINE BIOSCI