Eureka AIR delivers breakthrough ideas for toughest innovation challenges, trusted by R&D personnel around the world.

Isolated myeloid-like cell populations and methods of treatment therewith

A technology of cell populations and bone marrow cells, applied in the field of isolated mammalian cells, can solve problems such as undetermined cell numbers and unclear mechanisms

Inactive Publication Date: 2009-12-02
THE SCRIPPS RES INST
View PDF2 Cites 3 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although these cells have been used in several experimental models of angiogenesis, the mechanisms by which EPCs target neovasculature are not well understood, nor have strategies identified that effectively increase the number of cells contributing to specific vasculature

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Isolated myeloid-like cell populations and methods of treatment therewith
  • Isolated myeloid-like cell populations and methods of treatment therewith
  • Isolated myeloid-like cell populations and methods of treatment therewith

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0163] Example 1. Cell Separation and Enrichment; Mouse Lin - Preparation of HSC populations A and B.

[0164]general method. All in vivo evaluations were carried out in accordance with the NIH Guide for the Care and Use of Laboratory Animals (NIH Guide for the Care and Use of Laboratory Animals), and all evaluation methods were approved by The Scripps Research Institute (TSRI, La Jolla, CA). ) Animal Care and Use Committee (Animal Care and Use Committee) approval. Bone marrow cells were extracted from B6.129S7-Gtrosa26, Tie-2GFP, ACTbEGFP, FVB / NJ (rd / rd mice) or Balb / cBYJ adult mice (The Jackson Laboratory, ME).

[0165] then use A water-soluble Ficoll gradient (Sigma, St. Louis, MO), separated by density gradient, was used to isolate mononuclear cells, which were treated with biotin-conjugated lineage group antibodies (CD45, CD3, Ly-6G, CD11, TER-119, Pharmingen, San Diego, CA) labeled Lin for mice - filter. Using a magnetic separation device (AUTOMACS TM sorter, Mil...

Embodiment 2

[0171] Example 2. Intravitreal administration of cells in a murine model.

[0172] Eyelid clefts were cut on the eyelids of the mice with a fine blade to expose the eyeballs from P2 to P6. Then, using a No. 33 (Hamilton, Reno, NV) syringe with a needle, the lineage-negative HSC A population of the present invention (about 0.5 μl to about 1 μl of cell culture medium) was injected with about 10 5 cells) injected into the vitreous cavity.

Embodiment 3

[0173] Example 3. EPC transfection.

[0174] According to the manufacturer's protocol, use FuGENE TM 6 transfection reagent (Roche, Indianapolis, IN), mouse Lin - HSC (group A) also appends His with the T2 fragment encoding TrpRS 6 Tagged DNA (SEQ ID NO: 1, Figure 7 ) transfection. Will Lin - HSC cells (about 10 6 cells / ml) were suspended in OPTI- medium (Invitrogen, Carlsbad, CA). A mixture of DNA (about 1 μg) and FuGENE reagent (about 3 μl) was then added and the mixture was incubated at about 37° C. for about 18 hours. After incubation, cells were washed and harvested. The transfection efficiency of this system was approximately 17%, as confirmed by FACS analysis. Production of T2-TrpRS was confirmed by Western blotting. Add His 6 For the amino acid sequence of the T2-TrpRS tag, see Figure 8 SEQ ID NO: 2 in.

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

No PUM Login to View More

Abstract

The present invention provides an isolated myeloid-like cell population comprising a majority of cells that are lineage negative, and which express both CD44 antigen, CD11b antigen, and hypoxia inducible factor 1alpha (HIF-1alpha). These cells have beneficial vasculotrophic and neurotrophic activity when intraocularly administered to the eye of a mammal, particularly a mammal suffering from an ocular degenerative disease. The myeloid-like cells are isolated by treating bone marrow cells, peripheral blood cells or umbilical cord cells with an antibody against CD44 (hyaluronic acid receptor), against CD11b, CD14, CD33, or against a combination thereof and using flow cytometry to positively select CD44 and / or CD11b expressing cells therefrom. The isolated myeloid-like bone marrow cells of the invention can be transfected with a gene encoding a therapeutically useful protein, for delivering the gene to the retina.

Description

[0001] Cross References to Related Applications [0002] This application is a continuation-in-part of International Application Serial No. PCT / US2006 / 06411, filed February 24, 2006, which claims the benefit of U.S. Provisional Patent Application No. 60 / 656,037, filed February 24, 2005, so The above patent applications are incorporated herein by reference. [0003] Statement of Government Interests [0004] Portions of the research disclosed herein were supported by grant numbers EY11254, EY12598, EY13916, and EY14174 from the National Eye Institute of the National Institutes of Health. The United States Federal Government may have certain rights in this invention. field of invention [0005] The present invention relates to isolated mammalian cells. More specifically, the present invention relates to isolated cell populations that have the characteristics of myeloid cells and are capable of incorporating into the retinal vasculature when injected intravitreously into the e...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
Patent Type & Authority Applications(China)
IPC IPC(8): A01N63/00C12N5/06C12N5/08A61K35/12C12N5/074C12N5/0789
CPCA61K2035/124C12N5/0692C12N5/0647A61P9/00A61P9/10A61P27/02A61P27/06
Inventor M·弗里兰德M·R·里特S·K·莫尔诺V·马彻蒂
Owner THE SCRIPPS RES INST
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Eureka Blog
Learn More
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com