Isolated adult pluripotent stem cells and methods for isolating and cultivating thereof

• US 8,603,809 B2
• Filed: 04/08/2004
• Issued: 12/10/2013
• Est. Priority Date: 06/23/2003
• Status: Active Grant

• Current Assignee: Fraunhofer Gesellschaft Zur Foerderung Der Angewandten Forsching E.V.
• Original Assignee: Fraunhofer Gesellschaft Zur Foerderung Der Angewandten Forsching E.V.
• Inventors: Kruse, Charli
• Primary Examiner(s): Ton, Thaian N
• Application Number:
  US10/820,430
• Publication Number:
  US 20050014254A1
• Time in Patent Office: 3,533 Days
• Field of Search: 435/325, 435/378
• US Class Current: 435/325
• 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: A61K 35/39 Pancreas; Islets of Langerhans Langerhans cells of epidermis A61K35/36
• CPC Class Codes: C12N 5/0607 Non-embryonic pluripotent stem cells, e.g. MASC induced pluripotent stem cells C12N5/0696

Abstract

This invention relates to isolated pluripotent adult stem cells which are obtained from exocrine glandular tissue as well as methods of isolating and culturing these pluripotent stem cells.

Claims

1. A composition consisting of isolated pluripotent adult stem cells obtained from an exocrine glandular tissue of a salivary gland, a lacrimal gland, a sudoriferous gland, a sebaceous gland and/or gastrointestinal tissue, wherein the exocrine glandular tissue originates from a mammal, wherein the process for producing the isolated pluripotent adult stem cells comprises:

removal of exocrine glandular tissue;

the tissue thus removed is divided in such a gentle way that cell structures in resulting tissue fragments are largely preserved;

the divided tissue is cultured in a culture medium in the absence of additional specific growth factors or differentiation factors, whereby most of the differentiated cells become detached from the stem cells, whereupon the stem cells adhere on a bottom of a tissue culture vessel;

the remaining tissue and nonadherent differentiated cells are largely separated by a first change of medium and the remaining nonadherent cells are separated by additional changes of medium at intervals of about 2 to 3 days; and

isolating the pluripotent adult stem cells,

further wherein the isolated pluripotent adult stem cells are capable of differentiating into cell types of all three germ layers in a culture medium that does not contain any additional growth factors or differentiation factors after culturing under spatial conditions which ensure three dimensional contact of the cells.

2. The composition according to claim 1, wherein the exocrine glandular tissue is acinar tissue.

3. The composition according to claim 2, wherein the acinar tissue is derived from the pancreas, the parotid gland or the mandibular gland.

4. A stem cell culture consisting of the composition according to claim 1 in a culture medium adapted to allow stable maintenance and proliferation of the cells essentially without differentiation.

5. The stem cell culture according to claim 4, wherein the culture medium does not include any feeder cell layer.

13. The composition according to claim 1, wherein the mammal is a primate or a rodent.

14. The composition according to claim 1, wherein the mammal is a human.

15. The composition according to claim 1, wherein the mammal is a rat.

6. A primary stem cell culture obtained from exocrine glandular tissue from a salivary gland, a lacrimal gland, a sudoriferous gland, a sebaceous gland and/or gastrointestinal tissue of a mammal, wherein living cells from the exocrine glandular tissue present in the culture consist essentially of undifferentiated pluripotent adult stem cells.

7. The primary stem cell culture according to claim 6, wherein the living cells present in the culture consist of undifferentiated pluripotent adult stem cells.

8. The primary stem cell culture according to claim 6, wherein the exocrine glandular tissue is acinar tissue.

9. The primary stem cell culture according to claim 8, wherein the acinar tissue is derived from the pancreas, the parotid gland or the mandibular gland.

10. The primary stem cell culture according to claim 6, wherein the mammal is a primate or a rodent.

11. The primary stem cell culture according to claim 6, wherein the mammal is a human.

12. The primary stem cell culture according to claim 6, wherein the mammal is a rat.

16. A primary stem cell culture obtained from exocrine glandular tissue from a salivary gland, a lacrimal gland, a sudoriferous gland, a sebaceous gland and/or gastrointestinal tissue of a mammal, wherein living cells from the exocrine glandular tissue present in the culture consist essentially of undifferentiated adult stem cells which are capable of differentiating into cells of all three germ cell layers, wherein the process for producing the isolated pluripotent adult stem cells comprises:

removal of exocrine glandular tissue;

the tissue thus removed is divided in such a gentle way that cell structures in resulting tissue fragments are largely preserved;

the divided tissue is cultured in a culture medium in the absence of additional specific growth factors or differentiation factors, whereby most of the differentiated cells become detached from the stem cells, whereupon the stem cells adhere on a bottom of a tissue culture vessel;

the remaining tissue and nonadherent differentiated cells are largely separated by a first change of medium and the remaining nonadherent cells are separated by additional changes of medium at intervals of about 2 to 3 days; and

isolating the pluripotent adult stem cells,

further wherein the isolated pluripotent adult stem cells are capable of differentiating into cell types of all three germ layers in a culture medium that does not contain any additional growth factors or differentiation factors after culturing under spatial conditions which ensure three dimensional contact of the cells.

17. The primary stem cell culture according to claim 16, wherein the living cells present in the culture consist of undifferentiated adult stem cells which are capable of differentiating into cells of all three germ cell layers.

18. The primary stem cell culture according to claim 16, wherein the exocrine glandular tissue is acinar tissue.

19. The primary stem cell culture according to claim 18, wherein the acinar tissue is derived from the pancreas, the parotid gland or the mandibular gland.

20. The primary stem cell culture according to claim 16, wherein the mammal is a primate or a rodent.

21. The primary stem cell culture according to claim 16, wherein the mammal is a human.

22. The primary stem cell culture according to claim 16, wherein the mammal is a rat.

23. A composition consisting of isolated pluripotent adult stem cells obtained from an exocrine glandular tissue of a salivary gland, a lacrimal gland, a sudoriferous gland, a sebaceous gland and/or gastrointestinal tissue, wherein the exocrine glandular tissue originates from a mammal, wherein the process for producing the isolated pluripotent adult stem cells comprises:

removal of exocrine glandular tissue;

the tissue thus removed is divided in such a gentle way that cell structures in resulting tissue fragments are largely preserved;

the divided tissue is cultured in an incubation medium in the absence of specific growth factors or differentiation factors, whereby most of the differentiated cells become detached from the stem cells, whereupon the stem cells adhere on a bottom of a tissue culture vessel;

the remaining tissue and nonadherent differentiated cells are largely separated by a first change of medium and the remaining nonadherent cells are separated by additional changes of medium at intervals of about 2 to 3 days; and

isolating the pluripotent adult stem cells,

further wherein the isolated pluripotent adult stem cells are capable of differentiating into cell types of all three germ layers in a culture medium that does not contain any additional growth factors or differentiation factors after culturing under spatial conditions which ensure three dimensional contact of the cells.

24. A primary stem cell culture obtained from exocrine glandular tissue from a salivary gland, a lacrimal gland, a sudoriferous gland, a sebaceous gland and/or gastrointestinal tissue of a mammal, wherein living cells from the exocrine glandular tissue present in the culture consist essentially of undifferentiated adult stem cells which are capable of differentiating into cells of all three germ cell layers, wherein the process for producing the isolated pluripotent adult stem cells comprises:

removal of exocrine glandular tissue;

the tissue thus removed is divided in such a gentle way that cell structures in resulting tissue fragments are largely preserved;

the divided tissue is cultured in an incubation medium in the absence of specific growth factors or differentiation factors, whereby most of the differentiated cells become detached from the stem cells, whereupon the stem cells adhere on a bottom of a tissue culture vessel;

the remaining tissue and nonadherent differentiated cells are largely separated by a first change of medium and the remaining nonadherent cells are separated by additional changes of medium at intervals of about 2 to 3 days; and

isolating the pluripotent adult stem cells,

further wherein the isolated pluripotent adult stem cells are capable of differentiating into cell types of all three germ layers in a culture medium that does not contain any additional growth factors or differentiation factors after culturing under spatial conditions which ensure three dimensional contact of the cells.

25. A composition consisting of isolated pluripotent adult stem cells obtained from an exocrine glandular tissue of a salivary gland, a lacrimal gland, a sudoriferous gland, a sebaceous gland and/or gastrointestinal tissue, wherein the exocrine glandular tissue originates from a mammal, wherein the process for producing the isolated pluripotent adult stem cells comprises:

removal of exocrine glandular tissue;

the tissue thus removed is divided in such a gentle way that cell structures in resulting tissue fragments are largely preserved;

the divided tissue is cultured in an incubation medium plus serum in the absence of additional specific growth factors or differentiation factors, whereby most of the differentiated cells become detached from the stem cells, whereupon the stem cells adhere on a bottom of a tissue culture vessel;

the remaining tissue and nonadherent differentiated cells are largely separated by a first change of medium and the remaining nonadherent cells are separated by additional changes of medium at intervals of about 2 to 3 days; and

isolating the pluripotent adult stem cells,

further wherein the isolated pluripotent adult stem cells are capable of differentiating into cell types of all three germ layers in a culture medium that does not contain any additional growth factors or differentiation factors after culturing under spatial conditions which ensure three dimensional contact of the cells.

26. A primary stem cell culture obtained from exocrine glandular tissue from a salivary gland, a lacrimal gland, a sudoriferous gland, a sebaceous gland and/or gastrointestinal tissue of a mammal, wherein living cells from the exocrine glandular tissue present in the culture consist essentially of undifferentiated adult stem cells which are capable of differentiating into cells of all three germ cell layers, wherein the process for producing the isolated pluripotent adult stem cells comprises:

removal of exocrine glandular tissue;

the tissue thus removed is divided in such a gentle way that cell structures in resulting tissue fragments are largely preserved;

the divided tissue is cultured in an incubation medium plus serum in the absence of additional specific growth factors or differentiation factors, whereby most of the differentiated cells become detached from the stem cells, whereupon the stem cells adhere on a bottom of a tissue culture vessel;

the remaining tissue and nonadherent differentiated cells are largely separated by a first change of medium and the remaining nonadherent cells are separated by additional changes of medium at intervals of about 2 to 3 days; and

isolating the pluripotent adult stem cells,

further wherein the isolated pluripotent adult stem cells are capable of differentiating into cell types of all three germ layers in a culture medium that does not contain any additional growth factors or differentiation factors after culturing under spatial conditions which ensure three dimensional contact of the cells.