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

• US 9,206,393 B2
• Filed: 10/31/2013
• Issued: 12/08/2015
• 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 Forderung Der Angewandten Forschung EV
• Inventors: Kruse, Charli
• Primary Examiner(s): Ton, Thaian N
• Application Number:
  US14/068,681
• Publication Number:
  US 20140212965A1
• Time in Patent Office: 768 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: 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 method of producing adult pluripotent stem cells from mammalian exocrine glandular tissue, wherein said exocrine glandular tissue is isolated from a salivary gland, lacrimal gland, sudoriferous gland or sebaceous gland, and wherein said pluripotent adult stem cells are—

after culturing under spatial conditions which ensure three-dimensional contact of the cells—

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, the method comprising the following steps;

the exocrine glandular tissue is mechanically and enzymatically divided in such a manner that the cell structures in the resulting tissue fragments are largely preserved;

the divided tissue is cultured in an adherent culture;

the adherent cells persisting in the culture are further cultured and nonadherent differentiated cells are separated by a change of medium; and

adult pluripotent stem cells are obtained from the culture.

2. The method according to claim 1, characterized in that the divided tissue is first cultured under suitable conditions in tissue culture vessels, whereby most of the differentiated cells rapidly die in the course of a few days and become detached from the stem cells, whereupon the stem cells adhere on the bottom of the tissue culture vessel, and 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 a few days, preferably about 2 to 3 days.

3. The method according to claim 1, wherein the mammal is a primate.

4. The method according to claim 3, wherein the primate is a human.

5. The method according to claim 1, characterized in that the exocrine glandular tissue is acinar tissue.

6. The method according to claim 5, characterized in that the acinar tissue is isolated from the parotid gland or the mandibular gland.