Isolation and use of pluripotent stem cell population from adult neural crest-derived tissues

Patent metadata

Field	Value
Publication number	US-10106773-B2
Application number	US-201314382287-A
Country	US
Kind code	B2
Filing date	Mar 1, 2013
Priority date	Mar 1, 2012
Publication date	Oct 23, 2018
Grant date	Oct 23, 2018

Abstract

The present invention relates to methods of isolating a substantially homogeneous population of pluripotent stem cells from adult neural crest tissue (e.g., periodontal ligament) as well as pharmaceutical compositions comprising such isolated pluripotent stem cells. Methods of inducing the isolated pluripotent stem cells into specific cell lineages, such as neurogenic and retinogenic lineages, are also described. The isolated pluripotent stem cells find use in various regenerative medicine applications and the treatment of degenerative diseases.

First claim

The invention claimed is:

1. A method of isolating pluripotent stem cells comprising:
extracting cells from tissue derived from the neural crest, wherein the tissue derived from the neural crest is adult periodontal ligament or dental pulp from exfoliated deciduous teeth;
culturing said extracted cells under adherent conditions; and
isolating said cultured cells that express Connexin-43, wherein said isolating is
performed using a reagent with an affinity for said Connexin-43 to obtain a population of said cultured cells that express Connexin-43, wherein said population comprises at least 80% pluripotent stem cells.

2. The method of claim 1 , further comprising screening said extracted cells for expression of Connexin-43, at least one stem cell marker, or at least one neural crest marker prior to isolation.

3. The method of claim 2 , wherein said at least one stem cell marker or neural crest marker is Oct4, Nanog, Sox2, Klf4, p75 neurotrophin receptor, Nestin, Sox10, N-Cadherin, Notch1, BMP2, Snail, Slug, or combinations thereof.

4. The method of claim 1 , wherein said isolated stem cells are capable of differentiating into cells of the ectoderm, endoderm, and mesoderm lineages and generating teratomas when injected into immunodeficient mice.

5. A method of repairing damaged tissue in a subject in need thereof comprising administering to said subject pluripotent stem cells isolated using the method of claim 1 .

6. The method of claim 5 , wherein the stem cells are autologous.

7. The method of claim 5 , wherein said damaged tissue is neural tissue, retinal tissue, skeletal muscle tissue, cardiac tissue, bone or cartilage.

Assignees

• Univ Miami

Inventors

• Cheung Herman S
• Pelaez Daniel
• Huang C-Y Charles

Classifications

• C12N2501/115

  Basic fibroblast growth factor (bFGF, FGF-2) · CPC title

• C12N2501/415

  Wnt; Frizzeled · CPC title

• C12N2501/155

  Bone morphogenic proteins [BMP]; Osteogenins; Osteogenic factor; Bone inducing factor · CPC title

• C12N2501/11

  Epidermal growth factor [EGF] · CPC title

• C12N5/0607 Primary

  Non-embryonic pluripotent stem cells, e.g. MASC (induced pluripotent stem cells C12N5/0696) · CPC title