Pluripotent embryonic-like stem cells derived from teeth and uses thereof

• US 20050106724A1
• Filed: 02/05/2003
• Published: 05/19/2005
• Est. Priority Date: 02/06/2002
• Status: Abandoned Application

• Current Assignee: Stiftung Caesar
• Original Assignee: Stiftung Caesar
• Inventors: Hoffman, Karl-Heiz, Morsczexk, Christian, Schierholz, Joerg, Brenner, Norbert, Zeilhofer, Hans-Florian
• Application Number:
  US10/497,206
• Publication Number:
  US 20050106724A1
• Time in Patent Office: Days
• US Class Current: 435/366
• 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 1/02 Stomatological preparations, e.g. drugs for caries, aphtae, periodontitis
• CPC Class Codes: A61P 17/00 Drugs for dermatological disorders
• CPC Class Codes: A61P 19/00 Drugs for skeletal disorders
• CPC Class Codes: A61P 21/00 Drugs for disorders of the muscular or neuromuscular system
• CPC Class Codes: A61P 25/00 Drugs for disorders of the nervous system
• CPC Class Codes: A61P 43/00 Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• CPC Class Codes: A61P 9/00 Drugs for disorders of the cardiovascular system
• CPC Class Codes: C12N 2500/38 Vitamins
• CPC Class Codes: C12N 2501/39 Steroid hormones
• CPC Class Codes: C12N 2503/02 Drug screening
• CPC Class Codes: C12N 2510/00 Genetically modified cells
• CPC Class Codes: C12N 2533/00 Supports or coatings for cell culture, characterised by material
• CPC Class Codes: C12N 2533/40 Polyhydroxyacids, e.g. polymers of glycolic or lactic acid (PGA, PLA, PLGA); Bioresorbable polymers
• CPC Class Codes: C12N 5/0607 Non-embryonic pluripotent stem cells, e.g. MASC induced pluripotent stem cells C12N5/0696
• CPC Class Codes: C12N 5/0664 Dental pulp stem cells, Dental follicle stem cells
• CPC Class Codes: G01N 33/5008 for testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics
• CPC Class Codes: G01N 33/5017 for testing neoplastic activity
• CPC Class Codes: G01N 33/502 for testing non-proliferative effects
• CPC Class Codes: G01N 33/5044 involving specific cell types
• CPC Class Codes: G01N 33/5073 : Stem cells

Abstract

A stem cell which is obtainable from the non-embryonic tissue isolated from the dental follicle of tooth or wisdom tooth which are able to differentiate into a periodontal ligament like membrane structure.

Claims

1. A stem cell which is obtainable from the non-embryonic tissue isolated from the dental follicle of tooth or wisdom tooth which is capable of self-renewal and able to differentiate in vitro.

2. The stem cell of claim 1 capable of differentiation into cells of endodermal, ectodermal and mesodermal lineages.

3. The stem cell according to claim 1, which is capable of differentiating into a periodontal ligament like membrance structure.

4. The stem cell according to claim 1, which is able to differentiate into a mineralised tissue like structure.

5. The stem cell according to claim 1 which is a human cell.

6. A mesenchymal stem cell obtainable from the stem cell of claim 1.

7. A culture comprising:

a) stem cells according to claim 1,

b) a medium capable of supporting the proliferation of said stem cells,

c) a medium capable of supporting the differentiation of said stem cells into cells building a biological membrane with blood vessel or other vascular vessels.

8. The culture of claim 7, further comprising a proliferation factor or lineage commitment factor.

9. A method of isolating a stem cell according to claim 1, comprising the steps of

obtaining cells from a dental sac (dental follicle)

culturing the cells

10. The method according to claim 9 for isolating a clonal stem cell line, comprising the steps of

obtaining clonogenic cells from a dental follicle

culturing the cells in different media

diluting said cultured cells to clonal density;

culturing said diluted cells;

propagating those cultures having a single cell.

11. A clonal stem cell line obtainable by the method of claim 10.

12. The stem cell of claim 1 genetically engineered to express a gene or protein of interest.

13. A method of producing a genetically engineered stem cell that expresses a gene or protein of interest comprising the steps of

(a) transfecting the stem cell according to claim 1 with a DNA construct comprising at least one of a marker gene or a gene of interest;

(b) selecting for expression of the marker gene or gene of interest in the stem cells;

(c) culturing the stem cells selected in (b).

14. A genetically engineered stem cell obtainable by the method of claim 13.

15. A method for detecting the presence or activity of an agent which is a lineage-commitment factor comprising the steps of

contacting the stem cells of claim 1 with a sample suspected of containing an agent which is a lineage-commitment factor;

and

determining the lineage of the so contacted cells by mRNA expression, Antigen expression or other means;

wherein the lineage of the contacted cells indicates the presence or activity of a lineage-commitment factor in said sample.

16. A method of testing the ability of an agent, compound or factor to modulate the lineage-commitment of a lineage uncommitted cell which comprises

culturing the stem cells of claim 1 in a growth medium which maintains the stem cells as lineage uncommitted cells-,

adding the agent, compound or factor under test;

and

determining the lineage of the so contacted cells by mRNA expression, antigen expression or other means.

17. An assay system for screening agents, compounds or factors for the ability to modulate the lineage-commitment of a lineage uncommitted cell, comprising:

culturing the stem cells of claim 1 in a growth medium which maintains the stem cells as lineage uncommitted cells;

adding the agent, compound or factor under test; and

determining the lineage of the so contacted cells by mRNA expression, antigen expression or other means.

18. A method for detecting the presence or activity of an agent which is a proliferation factor comprising the steps of

contacting the stem cells of claim 1 with a sample suspected of containing an agent which is a proliferation factor;

determining the proliferation and lineage of the so contacted cells by mRNA expression, antigen expression or other means;

wherein the proliferation of the contacted cells without lineage commitment indicates the presence or activity of a proliferation factor in said sample.

19. A method of testing the ability of an agent, compound or factor to modulate the proliferation of a lineage uncommitted cell which comprises

culturing the stem cells of claim 1 in a growth medium which maintains the stem cells as lineage uncommitted cells;

adding the agent, compound or factor under test;

determining the proliferation and lineage of the so contacted cells by mRNA expression, antigen expression or other means.

20. A method for culturing stem cells of claim 1 into membranes which will be used for regenerating parodontitis, tendons, ligaments, fascia and others kinds of connecting tissues supporting differentiation to mesothelic/endothelic cells into blood vessels or ectodermal or into neuronal tissue.

21. A method for culturing stem cells of claim 1 in any media containing one substance for differentiation selected from the group of insulin, retinoic acid, indomethacin, 1-methyl-3-isobutylxanthine theophylline, transforming-growth-factor-beta (any), bone morphogenetic protein (any), Fibroblast growth factor (any), Epidermal Growth factor (any), Platelet derived growth factor (any), Vascular endothelial growth factor (any), hepatocyte growth factor, Interferon (any), Insulin like growth factor (any), Interleukine (any), nerve growth factor, or a combination thereof.

22. A method of ex vivo administration of a protein or gene of interest comprising the step of transfecting the stem cell of claim 1 with a vector comprising DNA or RNA which expresses a protein or gene of interest.

23. Use of the of stem cells according to claim 1, or cells or tissues derived there from, for the manufacturing of a medicament for preventing and/or treating cellular debilitations, derangements and/or dysfunctions and/or other disease states in mammals.

24. Use of the of stem cells according to claim 1, or cells or tissues derived there from, for the manufacturing of a medicament for tissue repair or transplantation in mammals.

25. Use of a mesodermal lineage committed cell derived from the stem cell of claim 1 for the manufacturing of a medicament for the prevention of and/or treatment of cellular debilitations, derangements and/or dysfunctions and/or for transplantations and/or other disease states in mammals.

26. Use of a mesodermal lineage committed cell derived from the stem cell of claim 1 for the manufacturing of a medicament for tissue repair or transplantation in mammals.

27. A pharmaceutical composition for the treatment of cellular debilitation, derangement and/or dysfunction in mammals, comprising:

a therapeutically effective amount of stem cells cells according to claim 1, or cells or tissues derived therefrom; and

a pharmaceutically acceptable medium or carrier.

28. The pharmaceutical composition of claim 27 further comprising a proliferation factor or lineage-commitment factor.

29. A scaffold serving as an extracellular matrix, consisting of polymers selected from the group of biodegradable polymers, non-biodegradable polymers and synthetic polymers to engineer teeth, to build teeth-surroundings bones/bone matrix alveolar bone or jaw bone, or to build arterial and venous vessels of the mouth by culturing the stem cells of claim 1 on matrices.

30. The scafford according to claim 29, whereby the scaffold matrix has cell binding domain sequences such as cadherins, CAMS or integrins.

31. A method of culturing of cells of claim 1 anyone of claims 1 wherein the cells are seeded on matrices in a bioreactor.