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Isolated monocyte populations and related therapeutic applications

A mononuclear cell and cell group technology, applied in the treatment of ocular vascular diseases, treatment or improvement of subjects' eye diseases, can solve problems such as no effective treatment methods

Inactive Publication Date: 2012-01-11
THE SCRIPPS RES INST
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although significant progress has been made in identifying factors that promote and inhibit angiogenesis, there are still no effective treatments for slowing or reversing the progression of these retinal degenerative diseases

Method used

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  • Isolated monocyte populations and related therapeutic applications
  • Isolated monocyte populations and related therapeutic applications
  • Isolated monocyte populations and related therapeutic applications

Examples

Experimental program
Comparison scheme
Effect test

example 1

[0082] Example 1, Isolation of a Monocyte Population

[0083] Peripheral blood or bone marrow can be used as source material for the procedures described here. As an example, we chose peripheral blood as the cell source due to its relative abundance of monocytes compared to bone marrow and the ease / safety of collection. For therapeutic use, it is desired that the cells do not have any bound compounds associated with selection. With this goal in mind, we have conceived and implemented methods to differentiate monocytes from other mononuclear cells such as lymphocytes based solely on physical properties such as size, granularity and density. The first method we have developed is based on FACS, without the use of antibodies, and sensitively separates monocytes from lymphocytes based on differences in cell size and granularity. Results showing monocyte populations isolated by this method are presented in figure 1 In A. Red blood cells and granulocytes present in whole blood ar...

example 2

[0090] Example 2, Treatment of Ocular Vascular Disease with Isolated Monocyte Populations

[0091] The therapeutic activity of monocyte populations isolated using the methods described herein was tested using a mouse model of oxygen-induced retinopathy. Oxygen-induced retinopathy mice were generated as described in Ritter et al., J. Clin. Invest. 116:3266-76, 2006. Specifically, C57BL / 6J mice were induced to develop oxygen-induced retinopathy according to the protocol described in Smith et al., Invest. Ophthalmol. Vis. Sci. 35: 101-111, 1994. As a comparison, BALB / cByJ mice were also treated with the same conditions. Briefly, P7 pups and their mothers were transferred from a room air environment to an environment containing 75% oxygen for 5 days and then returned to the room air environment. Create and maintain a hyperoxic environment using a BioSpherix chamber. Under these conditions, during hyperoxia in C57BL / 6J mice, a large hypovascular zone forms in the central retina,...

example 3

[0093]Example 3, Additional Properties and Activities of Isolated Monocyte Populations

[0094] In order to demonstrate that the cells we isolated were different from known cell populations for clinical use or development, we have labeled peripheral blood samples for the expression of aldehyde dehydrogenase, when expressed at high levels (ALDH br ), which discriminates CD34 + cells, CD133 + cell, kit + Cell, lineage antigen negative (Lin - )cell. We found that peripheral blood samples ( image 3 ) essentially lacks such markers, matching the notion that stem cells are expected to be extremely rare in immobile peripheral blood.

[0095] CD34 is a marker of hematopoietic stem cells and has been used to select cells for various clinical applications. We have found that such cells may include or adversely affect the outcome of the therapeutic applications described herein. Specifically, to determine the behavior of undifferentiated stem cells after intraocular injection, we...

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PUM

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Abstract

The invention provides methods of using isolated monocyte populations to treat subjects suffering from various ocular vascular disease or ocular degenerative disorders. The present invention also provides novel methods for isolating substantially pure monocyte populations. The methods involve extracting a blood sample or a bone marrow sample from a subject, debulking red blood cells from the sample, and then separating remaining red blood cells and other cell types in the sample from monocytes. Instead of using any selection or labeling agents, the red blood cells and other cell types are separated from monocytes based on their size, granularity or density. The isolated monocytes can be further activated in vitro or ex vivo prior to being administered to a subject. Isolated cell populations containing substantially pure CD14+ / CD33+ monocytes are also provided in the invention.

Description

[0001] statement of government support [0002] This invention was made with support in part by the US Government under Grant Nos. EY1254, EY14174, and EY017540 by the National Institutes of Health. Accordingly, the US Government has certain rights in this invention. [0003] Cross References to Related Applications [0004] The subject patent application claims the benefit of priority to US Provisional Patent Applications Serial Nos. 61 / 208,173 (filed February 20, 2009) and 61 / 283,244 (filed November 30, 2009), respectively. The entire disclosures of these prior applications are hereby incorporated by reference in their entirety and for all purposes. Background technique [0005] Ocular vascular diseases such as age-related macular degeneration (ARMD) and diabetic retinopathy (DR) are attributed to abnormal neovascularization of the abnormal choroid or retina, respectively. They are the leading cause of vision loss in industrialized countries. Since the retina is composed...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/0786A01N63/02A61K35/12
CPCC12N2501/052C12N5/0645C12N2501/21G01N2015/1497A61K2035/124C12N2501/051G01N2015/149G01N15/042A61P27/02A61P27/06A61P9/10G01N15/149A61K39/4614A61K39/464A61K39/4622A61K2239/31A61K35/12
Inventor 马丁·弗里德伦德马修·R·里特尔史黛丝·K·莫雷诺穆罕默德·A·埃尔-卡拉
Owner THE SCRIPPS RES INST
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