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WO2011144901A4 - Expansion and directed differentiation of epidermal neural crest stem cells - Google Patents
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WO2011144901A4 - Expansion and directed differentiation of epidermal neural crest stem cells - Google Patents

Expansion and directed differentiation of epidermal neural crest stem cells Download PDF

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Publication number
WO2011144901A4
WO2011144901A4 PCT/GB2011/000763 GB2011000763W WO2011144901A4 WO 2011144901 A4 WO2011144901 A4 WO 2011144901A4 GB 2011000763 W GB2011000763 W GB 2011000763W WO 2011144901 A4 WO2011144901 A4 WO 2011144901A4
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stem cells
neural crest
crest stem
inducing
cells
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Ceased
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PCT/GB2011/000763
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French (fr)
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WO2011144901A1 (en
Inventor
Maya Sieber-Blum
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Newcastle University of Upon Tyne
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Newcastle University of Upon Tyne
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Priority claimed from GBGB1008440.8A external-priority patent/GB201008440D0/en
Priority claimed from GBGB1013895.6A external-priority patent/GB201013895D0/en
Priority claimed from GBGB1020251.3A external-priority patent/GB201020251D0/en
Application filed by Newcastle University of Upon Tyne filed Critical Newcastle University of Upon Tyne
Publication of WO2011144901A1 publication Critical patent/WO2011144901A1/en
Publication of WO2011144901A4 publication Critical patent/WO2011144901A4/en
Anticipated expiration legal-status Critical
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    • C12N5/06—Animal cells or tissues; Human cells or tissues
    • C12N5/0602—Vertebrate cells
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    • C12N5/00—Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor
    • C12N5/06—Animal cells or tissues; Human cells or tissues
    • C12N5/0602—Vertebrate cells
    • C12N5/0618—Cells of the nervous system
    • C12N5/062—Sensory transducers, e.g. photoreceptors; Sensory neurons, e.g. for hearing, taste, smell, pH, touch, temperature, pain
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    • C12N5/06—Animal cells or tissues; Human cells or tissues
    • C12N5/0602—Vertebrate cells
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    • C12N5/06—Animal cells or tissues; Human cells or tissues
    • C12N5/0602—Vertebrate cells
    • C12N5/0625—Epidermal cells, skin cells; Cells of the oral mucosa
    • C12N5/0626—Melanocytes
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    • C12N5/0602—Vertebrate cells
    • C12N5/0652—Cells of skeletal and connective tissues; Mesenchyme
    • C12N5/0654—Osteocytes, Osteoblasts, Odontocytes; Bones, Teeth
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    • C12N2501/00—Active agents used in cell culture processes, e.g. differentation
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    • C12N2501/00—Active agents used in cell culture processes, e.g. differentation
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    • C12N2501/00—Active agents used in cell culture processes, e.g. differentation
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    • C12N2501/00—Active agents used in cell culture processes, e.g. differentation
    • C12N2501/10—Growth factors
    • C12N2501/125—Stem cell factor [SCF], c-kit ligand [KL]
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    • C12N2501/10—Growth factors
    • C12N2501/13—Nerve growth factor [NGF]; Brain-derived neurotrophic factor [BDNF]; Cilliary neurotrophic factor [CNTF]; Glial-derived neurotrophic factor [GDNF]; Neurotrophins [NT]; Neuregulins
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    • C12N2501/00—Active agents used in cell culture processes, e.g. differentation
    • C12N2501/40—Regulators of development
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    • C12N2506/00—Differentiation of animal cells from one lineage to another; Differentiation of pluripotent cells
    • C12N2506/03—Differentiation of animal cells from one lineage to another; Differentiation of pluripotent cells from non-embryonic pluripotent stem cells

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  • Health & Medical Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Biomedical Technology (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Zoology (AREA)
  • Organic Chemistry (AREA)
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  • Bioinformatics & Cheminformatics (AREA)
  • Genetics & Genomics (AREA)
  • Wood Science & Technology (AREA)
  • Cell Biology (AREA)
  • Neurology (AREA)
  • Biochemistry (AREA)
  • General Engineering & Computer Science (AREA)
  • General Health & Medical Sciences (AREA)
  • Microbiology (AREA)
  • Neurosurgery (AREA)
  • Developmental Biology & Embryology (AREA)
  • Dermatology (AREA)
  • Physics & Mathematics (AREA)
  • Acoustics & Sound (AREA)
  • Analytical Chemistry (AREA)
  • Orthopedic Medicine & Surgery (AREA)
  • Rheumatology (AREA)
  • Medicines Containing Material From Animals Or Micro-Organisms (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)

Abstract

The invention relates to the directed differentiation of epidermal neural crest stem cells, and more specifically human epidermal neural crest stem cells (hEPI-NCSC), for example to their differentiation into dopaminergic neurons, dopaminergic neuronal progenitors, sensory neurons, sympathetic neurons, cholinergic neurons, melanocytes, Schwann cells, smooth muscle, osteogenic differentiation into osteocytes, bone and cartilage and adrenergic differentiation.

Claims

AMENDED CLAIMS received by the International Bureau on 12 December 2011 (12.12.2011 ) Amended Claims (Art 19 PCT)
1. A method for inducing or stimulating differentiation of a population of EPI- NSCS cells into dopaminergic neurons or dopaminergic neuronal progenitors which includes the steps of:
I. Culturing said population of epidermal neural crest stem cells (EPI- NCSCs) in the presence of a neural progenitor (NP) medium selected to induce or stimulate progression of EPI-NCSCs to a neural stem cell like state; and
II. Culturing said population of cells in the presence of a patterning factor (PF) medium selected to induce or stimulate differentiation of the cells into dopaminergic neurons or dopaminergic neuronal progenitors.
2. A method for inducing or stimulating differentiation of epidermal neural crest stem cells as in Claim 1 wherein in step I. the induced or stimulated progression of EPI-NCSCs to a neural stem cell like state results in the cells changing morphology from stellate morphology to a more elongated shape with often elaborate short processes.
3. A method for inducing or stimulating differentiation of epidermal neural crest stem cells as in Claims 1 or 2 wherein in step II. the induced or stimulated stimulate differentiation of the cells into dopaminergic neurons or dopaminergic neuronal progenitors results in the cells becoming confluent and changing morphology by elaborating long processes and the soma being first rounded then assuming shape of dopaminergic neurons.
4. A method for inducing or stimulating differentiation of epidermal neural crest stem cells as in any of the previous Claims wherein the epidermal neural crest stem cells are a substantially pure population of epidermal neural crest stem cells.
5. A method for inducing or stimulating differentiation of epidermal neural crest stem cells as in any of the previous Claims wherein the method is for inducing or stimulating differentiation of human epidermal neural crest stem cells.
6. A method for inducing or stimulating differentiation of epidermal neural crest stem cells as in any of the previous Claims wherein the method includes an expansion step expanding the population of epidermal neural crest stem cells (EPI-NCSCs) prior to step I.
7. A method for inducing or stimulating differentiation of epidermal neural crest stem cells as in Claim 6 wherein the expansion step comprises culturing the isolated epidermal neural crest stem cells in expansion media.
8. A method for inducing or stimulating differentiation of epidermal neural crest stem cells as in Claim 7 wherein expansion media comprises;
proliferation media, FGF2, EGF, FBS, ITS+3, GlutaMAX ®, Amphotericin, Penicillin/Streptomycin.
9. A method for inducing or stimulating differentiation of epidermal neural crest stem cells as in any of the previous Claims wherein the Neural
Progenitor (NP) medium comprises;
FGF-2
EGF SCF NT-3
10. A method for inducing or stimulating differentiation of epidermal neural crest stem cells as in any of the previous Claims wherein the patterning factor (PF) medium comprises;
76 SHH ["sonic hedgehog"]
FGF-8
TGF-p2
GDNF [glial derived neurotrophic factor]
11.A method for inducing or stimulating differentiation of epidermal neural crest stem cells as in Claim 10 wherein the patterning factor (PF) medium further comprises;
db c-AMP
combinations of neurotrophic factors (NGF, BDNF, NT-3)
12. A method for inducing or stimulating differentiation of epidermal neural crest stem cells as in any of the previous Claims wherein the epidermal neural crest stem cells are obtained from the bulge of a hair follicle obtained from a subject.
13. A method for inducing or stimulating differentiation of epidermal neural crest stem cells as in Claim 12 wherein the subject is mammalian.
14. A method for inducing or stimulating differentiation of epidermal neural crest stem cells as in Claims 12 or 13 wherein the subject is human.
15. A method for inducing or stimulating differentiation of epidermal neural crest stem cells as in any of previous Claims 12 to 14 wherein the method further comprises the step of returning the differentiated cells into the subject.
16. A population of differentiated cells obtained by the method of any of
Claims 1 to 14.
17. Cells obtained by the method of any of Claims 1 to 14 for the treatment of a disease state.
77
18. Cells obtained by the method of any of Claims 1 to 14 for the treatment of Parkinson's disease.
19. Cells obtained by the method of any of Claims 1 to 14 for the treatment of degenerative diseases.
20. Human epidermal neural crest stem cells (hEPI-NCSC) for use in the treatment of any of the diseases referred to in Claims 17 to 19.
21. Dopaminergic neurons or dopaminergic neuronal progenitors obtained by the method of any of Claims 1 to 15.
22. A patterning factor (PF) media for the directed differentiation of hEPI- NCSCs into dopaminergic neurons or dopaminergic neuronal progenitors comprising;
SHH ["sonic hedgehog"]
FGF-8
TGF-P2
GDNF [glial derived neurotrophic factor]
23. A patterning factor (PF) media for the directed differentiation of hEPI- NCSCs as in Claim 22, further comprising
db c-AMP; and
combinations of neurotrophic factors (NGF, BDNF, NT-3)
24. A neural progenitor (NP) media for the progression of hEPI-NCSCs to a neural stem cell like state comprising;
FGF-2
EGF SCF NT-3
78
25. A method of treating Parkinson's disease comprising administering a therapeutically active amount of said dopaminergic neurons or
dopaminergic neuronal progenitors.
26. A method of treating Parkinson's disease comprising administering a
therapeutically active amount of human epidermal neural crest stem cells.
79
PCT/GB2011/000763 2010-05-20 2011-05-20 Expansion and directed differentiation of epidermal neural crest stem cells Ceased WO2011144901A1 (en)

Applications Claiming Priority (6)

Application Number Priority Date Filing Date Title
GBGB1008440.8A GB201008440D0 (en) 2010-05-20 2010-05-20 Directed differentiation of human epidermal neural crest stem cells
GB1008440.8 2010-05-20
GBGB1013895.6A GB201013895D0 (en) 2010-08-19 2010-08-19 Directed differentiation of human epidermal neural crest stem cells
GB1013895.6 2010-08-19
GB1020251.3 2010-11-30
GBGB1020251.3A GB201020251D0 (en) 2010-11-30 2010-11-30 Human epidermal neural crest stem cells (hEPI-NCSC) - Characterisation and directed differentiation into bone cells

Publications (2)

Publication Number Publication Date
WO2011144901A1 WO2011144901A1 (en) 2011-11-24
WO2011144901A4 true WO2011144901A4 (en) 2012-03-01

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PCT/GB2011/000763 Ceased WO2011144901A1 (en) 2010-05-20 2011-05-20 Expansion and directed differentiation of epidermal neural crest stem cells

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Cited By (1)

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US11564901B2 (en) 2007-01-31 2023-01-31 Biosuccess Biotech Co., Ltd. Compositions and methods of use of phorbol esters

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US9636317B2 (en) 2007-01-31 2017-05-02 Biosuccess Biotech Co. Ltd. Compositions and methods of use of phorbol esters
US9974764B2 (en) 2007-01-31 2018-05-22 Biosuccess Biotech Co. Ltd. Compositions and methods of use of phorbol esters in the treatment of neoplasms
US9533938B2 (en) 2007-01-31 2017-01-03 Biosuccess Biotech Co., Ltd. Compositions and methods of use of phorbol esters for the treatment of stroke
US10369222B2 (en) 2012-01-18 2019-08-06 Biosuccess Biotech Co., Ltd. Compositions and methods of use of phorbol esters for the treatment of stroke
EP2170053A4 (en) 2007-01-31 2010-06-23 Biosuccess Biotech Company COMPOSITIONS AND METHODS OF USING PHORBOL ESTERS
US10099996B2 (en) 2012-01-18 2018-10-16 Biosuccess Biotech Co. Ltd. Compositions and methods of use of phorbol esters in the treatment of neoplasms
US9550722B2 (en) 2012-01-18 2017-01-24 Biosuccess Biotech Co. Ltd. Compositions and methods of use of phorbal esters for the treatment of stroke
US20160040126A1 (en) * 2012-02-21 2016-02-11 Korea University Research And Business Foundation Regulation of differentiation into dopaminergic neurons by metalloprotease
MX2020005668A (en) 2017-11-30 2020-11-24 Univ Kyoto Method for culture of cells.
SG11202011395RA (en) * 2018-06-22 2020-12-30 Skin2Neuron Pty Ltd Expansion and differentiation of neuronal precursor cells
WO2020165615A1 (en) 2019-02-14 2020-08-20 Omnion Research International D.O.O. Method of transformation of hair folicle cells into neurons, method and scale for estimating risk for onset of a particular brain disease
CN111117968B (en) * 2019-07-30 2020-09-08 武汉赛尔朗灵科技有限公司 Method for establishing fluorescent nude mouse tumor model based on human brain glioma primary cells

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US7666675B2 (en) * 2001-09-20 2010-02-23 Anticancer, Inc. Nestin-expressing hair follicle stem cells
US20080057028A1 (en) 2002-10-02 2008-03-06 Institute Nationale De La Sante Et De La Recherche Medicale (Inserm) Vegf-C or Vegf-D Materials and Methods for Stimulation of Neural Stem cells
DE602004021509D1 (en) 2003-11-26 2009-07-23 Eisai R&D Man Co Ltd Specific marker Lmx1a for dopaminergic neurons
US8030072B2 (en) * 2005-03-15 2011-10-04 Newcastle University Method of isolating epidermal neural crest stem cells

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US11564901B2 (en) 2007-01-31 2023-01-31 Biosuccess Biotech Co., Ltd. Compositions and methods of use of phorbol esters

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