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Cells exhibiting neuronal cell progenitor characteristics and methods of making them

a cell and progenitor cell technology, applied in the field of cells exhibiting neuronal progenitor cell characteristics, can solve the problems of terminally differentiated neurons being significantly limited in their ability to proliferate, cns or peripheral nervous systems, and difficult to achieve treatment of cns or pns diseases that require the transplantation of terminally differentiated neurons

Inactive Publication Date: 2006-11-09
SANBIO
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

A limitation in the research and treatment of Central Nervous System (CNS) or Peripheral Nervous System (PNS) diseases is the conventional recognition that terminally differentiated neurons are significantly limited in their ability to proliferate.
Accordingly, any treatment of CNS or PNS diseases that requires transplant of terminally differentiated neurons is difficult to accomplish.
However, such CPCs are rare and difficult to isolate from donors.
However, the use of embryonic stem cells raises a number of ethical concerns, and so is a disfavored source of stem cells for production of CPCs.
Additionally, embryonic stem cells can be tumorigenic, which generates safety concerns as to any transplant procedure that could potentially result in the delivery of embryonic stem cells to a patient such as creation of a CPC graft from embryonic stem cells.
However, the early progenitor cells of Prockop may be undesirable for use because they seem to display a very immature neuronal phenotype whose clinical efficacy is not well understood.
Accordingly, there is a scarcity of conventionally available and suitable sources of CPCs for use, for example, in the research and treatment of CNS or PNS diseases.
Further, there is a scarcity of methods that can be used to produce such CPCs in a suitable manner suitable for use.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

example 1

[0052] Human MASCs (PT-2501, BioWhittaker, Walkersville, Md.) were allowed to grow in culture in alpha-MEM containing 10% FBS generally according to E. Sudbeck et al., Structure-based design of specific inhibitors of Janus kinase 3 as apoptosis-inducing antileukemic agents. Clin. Cancer Res. 5, 1569-1582 (1999). The MASCs were incubated with 40 ug / ml 4-(4′-hydroxyphenyl)amino-6,7-dimethoxyquinazoline (WHI-131, Calbiochem, San Diego, Calif.) for two days. The WHI-131 was washed off after 2 days.

example 2

[0053] Human MASCs, prepared according to the Materials and Methods section, are allowed to grow in culture in alpha-MEM containing 10% FBS generally according to E. Sudbeck et al., Structure-based design of specific inhibitors of Janus kinase 3 as apoptosis-inducing antileukemic agents. Clin. Cancer Res. 5, 1569-1582 (1999). Once the culture has reached 90% confluence, several RNAs, designed using the BLOCK-iT™ RNAi Designer (Invitrogen) are incubated with the culture for a period of time sufficient to silence Sox9 expression, using BLOCK-iT™ protocols available from Invitrogen. Resulting CPCs are isolated from untransdifferentiated MASC's by sequential selection using magnetic beads coated with appropriate antibodies such as anti-EfnB2 (positive selection for CPCs), anti-CD90 (negative selection for CPCs), and anti-PDGF receptor beta (negative selection for CPCs). The antibodies and coated beads may be obtained from commercial suppliers. The cells in PBS are incubated with coated ...

example 3

[0054] Human MASCs, prepared according to the Materials and Methods section, are allowed to grow in culture in alpha-MEM containing 10% FBS generally according to E. Sudbeck et al., Structure-based design of specific inhibitors of Janus kinase 3 as apoptosis-inducing antileukemic agents. Clin. Cancer Res. 5, 1569-1582 (1999). Antisense oligomers to Hes 1 are generated according to techniques disclosed in any one of H. Moulton et al., Peptide-assisted delivery of steric-blocking antisense oligomers. Curr Opin Mol Ther. 2003 April;5(2):123-32; C. Stein et al., Antisense oligonucleotides as therapeutic agents—is the bullet really magical? Science. 1993 Aug. 20;261(5124):1004-12; or C. Helene, The anti-gene strategy: control of gene expression by triplex-forming-oligonucleotides. Anticancer Drug Des. 1991 December;6(6):569-84. Once the MASC culture reaches 90% confluence, the Hes-1 antisense oligomers are incubated with the MASCs for a period sufficient to downregulate Hes-1 expression,...

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PUM

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Abstract

Disclosed are cells exhibiting neuronal progenitor cell characteristics, and methods of making them from marrow adherent stem cells by regulating cellular pathways in the marrow adherent stem cells that are associated with glial transdifferentiation of the marrow adherent stem cells.

Description

CROSS-REFERENCE TO RELATED APPLICATIONS [0001] This application claims benefit of U.S. provisional patent application 60 / 561,613, filed Apr. 12, 2004, which is incorporated herein by reference in its entiretyFIELD OF THE INVENTION [0002] The invention relates to cells exhibiting neuronal progenitor cell characteristics, and methods of making them from marrow adherent stem cells by regulating cellular pathways in the marrow adherent stem cells that are associated with glial transdifferentiation of the marrow adherent stem cells. BACKGROUND OF THE INVENTION [0003] A limitation in the research and treatment of Central Nervous System (CNS) or Peripheral Nervous System (PNS) diseases is the conventional recognition that terminally differentiated neurons are significantly limited in their ability to proliferate. Accordingly, any treatment of CNS or PNS diseases that requires transplant of terminally differentiated neurons is difficult to accomplish. [0004] One proposed approach to overcom...

Claims

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Application Information

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IPC IPC(8): A61K48/00C12N5/08A61K35/12C12N5/0793
CPCA61K35/12C12N5/0619C12N2501/01C12N2501/115C12N2506/1353C12N2501/155C12N2501/415C12N2501/60C12N2501/70C12N2501/13A61P25/00A61P25/02C12N5/0602A61K35/28C12N5/06C12N2501/727
Inventor DEZAWA, MARI
Owner SANBIO
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