Cells exhibiting neuronal progenitor cell characteristics and methods of making them

• US 9,441,199 B2
• Filed: 06/09/2015
• Issued: 09/13/2016
• Est. Priority Date: 04/12/2004
• Status: Active Grant

• Current Assignee: Sanbio, Inc. (SanBio Co., Ltd.)
• Original Assignee: Sanbio, Inc. (SanBio Co., Ltd.)
• Inventors: Dezawa, Mari
• Primary Examiner(s): Ton, Thaian N
• Application Number:
  US14/735,005
• Publication Number:
  US 20150267169A1
• Time in Patent Office: 462 Days
• US Class Current: 1/1
• CPC Class Codes: A61K 35/12 Materials from mammals; Compositions comprising non-specified tissues or cells; Compositions comprising non-embryonic stem cells; Genetically modified cells vaccines or medicinal preparations containing antigens or antibodies A61K39/00
• CPC Class Codes: A61P 25/00 Drugs for disorders of the nervous system
• CPC Class Codes: A61P 25/02 for peripheral neuropathies
• CPC Class Codes: C12N 2501/01 Modulators of cAMP or cGMP, e.g. non-hydrolysable analogs, phosphodiesterase inhibitors, cholera toxin
• CPC Class Codes: C12N 2501/115 Basic fibroblast growth factor (bFGF, FGF-2)
• CPC Class Codes: C12N 2501/13 Nerve growth factor [NGF]; Brain-derived neurotrophic factor [BDNF]; Cilliary neurotrophic factor [CNTF]; Glial-derived neurotrophic factor [GDNF]; Neurotrophins [NT]; Neuregulins
• CPC Class Codes: C12N 2501/155 Bone morphogenic proteins [BMP]; Osteogenins; Osteogenic factor; Bone inducing factor
• CPC Class Codes: C12N 2501/415 Wnt; Frizzeled
• CPC Class Codes: C12N 2501/60 Transcription factors
• CPC Class Codes: C12N 2501/70 Enzymes
• CPC Class Codes: C12N 2501/727 Kinases (EC 2.7.)
• CPC Class Codes: C12N 2506/1353 from bone marrow mesenchymal stem cells (BM-MSC)
• CPC Class Codes: C12N 5/0619 Neurons

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.

Claims

1. A method for making a population of mammalian mitotic cells that express nestin and EfnB2, wherein the method comprises culturing mammalian marrow adherent stem cells (MASCs) in the presence of a Janus kinase (JAK) inhibitor and isolating the mitotic cells that express nestin and EfnB2;

thereby providing a population of mammalian mitotic cells, in the absence of a JAK inhibitor, that express nestin and EfnB2.

2. The method of claim 1, wherein the mammalian MASCs are selected from the group consisting of human marrow adherent stem cells, rat marrow adherent stem cells, mouse marrow adherent stem cells, primate marrow adherent stem cells, pig marrow adherent stem cells, cow marrow adherent stem cells, and sheep marrow adherent stem cells.

3. The method of claim 2, wherein the mammalian MASCs are human MASCs.

4. The method of claim 1, wherein the JAK inhibitor is a synthetic small molecule organic compound.

5. The method of claim 4, wherein the small molecule JAK inhibitor is a dimethoxyquinazoline compound.

6. The method of claim 5, wherein the dimethoxyquinazoline compound is selected from the group consisting, of 4-(phenyl)-amino-6,7-dimethoxyquinazoline;

4-(4′

-hydroxyphenyl)-amino-6,7-dimethoxyquinazoline;

4-(3′

-bromo-4′

-hydroxylphenyl)-amino-6,7-dimethoxyquinazoline; and

4-(3′

,5′

-dibromo-4′

-hydroxylphenyl)-amino-6,7-dimethoxyquinazoline.

7. The method of claim 6, wherein the dimethoxyquinazoline compound is 4-(4′

-hydroxyphenyl)-amino-6,7-dimethoxyquinazoline.

8. The method of claim 1, wherein the JAK inhibitor is a cAMP elevating agent.

9. The method of claim 8, wherein the cAMP elevating agent is selected from the group consisting of forskolin and 3-isobutyl-1-methylxanthine (IBMX).

10. The method of claim 1, wherein the JAK inhibitor is selected from the group consisting of 4,5-dimethoxy-2-nitrobenzoic acid and 4,5-dimethoxy-2-nitrobenzamide.

11. The method of claim 1, wherein the JAK inhibitor is a RNAi.

12. The method of claim 1, wherein the JAK inhibitor is an antisense oligonucleotide that downregulates JAK activity.

13. The method of claim 1, wherein the JAK inhibitor is a polypeptide.

14. The method of claim 13, wherein the culturing comprises transfection of the mammalian MASCs with a polynucleotide encoding the polypeptide.

15. The method of claim 1, wherein subsequent culture of the mammalian mitotic cells that express nestin and EfnB2;

in the presence of forskolin, basic fibroblast growth factor and ciliary neurotrophic factor, and in the absence of the JAK inhibitor;

generates a population comprising mammalian MAP-2-positive cells.