Mammalian pluripotent stem cells, methods for their production, and uses thereof

Patent metadata

Field	Value
Publication number	US-2016030484-A1
Application number	US-201514816661-A
Country	US
Kind code	A1
Filing date	Aug 3, 2015
Priority date	Aug 4, 2014
Publication date	Feb 4, 2016
Grant date	—

Abstract

The disclosure relates to nerve derived adult pluripotent stem cells characterized by expression of Oct4, Sox2, c-Myc, and Klf4, methods for obtaining them, and their use.

First claim

1 . A mammalian nerve derived adult pluripotent stem cell (NEDAPS cell), characterized by expression of Oct4, Sox2, c-Myc, and Klf4.

2 . The NEDAPS cell of claim 1 which is isolated.

3 . The NEDAPS cell of claim 1 which is characterized by one, two, three, four or all of the following:
(a) capability of differentiating into a mesoderm cell type, optionally a mesenchymal cell type;
(b) capability of differentiating into an endoderm cell type;
(c) capability of differentiating into an ectoderm cell type, optionally a neural stem cell;
(d) motility; and
(e) adherence to glass or plastic substrate in culture.

4 . The NEDAPS cell of claim 3 wherein the mesenchymal cell type is (a) an osteoblast or (b) an endothelial cell.

5 . A population of NEDAPS cells according to claim 1 .

6 . The population of claim 5 which is isolated, optionally which is at least 55%, at least 60%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 99% homogeneous.

7 . A method of producing the population of claim 6 , comprising
(a) culturing a peripheral nerve exposed to NEDAP cell proliferation conditions ex vivo; or (b) culturing cells from a peripheral nerve exposed to NEDAP cell proliferation conditions in a subject in vivo, and
(i) optionally, harvesting the peripheral nerve prior to culturing; and
(ii) optionally, exposing the peripheral nerve to the NEDAP cell proliferation conditions prior to harvesting.

8 . The method of claim 7 , wherein the NEDAP cell proliferation conditions comprise:
(a) exposing the peripheral nerve to BMP2; (b) exposing the peripheral nerve to a neuroinflammatory agent other than BMP2; (c) applying trauma to the peripheral nerve; (d) a combination of two of (a)-(c); or (e) a combination of three of (a)-(c).

9 . The method of claim 8 , wherein the NEDAP cell proliferation conditions comprise exposing the peripheral nerve to BMP2 and wherein:
(a) the peripheral nerve is exposed to BMP2 in a subject in vivo, optionally wherein:
(i) BMP2 is applied directly to the peripheral nerve, optionally in an amount ranging from 10 ng to 1 milligram;
(ii) BMP2 is administered to the subject via intramuscular injection, optionally in an amount ranging from 10 ng to 1 milligram; or
(iii) the subject is exposed to conditions that result in local production of BMP2, optionally wherein the conditions comprise a bone fracture, blunt injury, thermal injury, or electric shock; or
(b) the peripheral nerve is exposed to BMP2 ex vivo by culturing the nerve in a medium comprising BMP2, optionally in a concentration ranging from 5 ng/ml to 1 mg/ml.

10 . The method of claim 8 , wherein the NEDAP cell proliferation conditions comprise applying trauma to the peripheral nerve, optionally wherein the trauma is mechanical trauma, electrical stimulation, an ultrasonic shock wave, or a thermal insult.

11 . The method of claim 8 , wherein the NEDAP cell proliferation conditions comprise exposing the peripheral nerve to a neuroinflammatory agent other than BMP2, optionally wherein the neuroinflammatory agent is tumor necrosis factor alpha, Interleukin-1 Beta, nerve growth factor, histamine, Interleukin 6, or a combination thereof.

12 . The method of claim 7 , wherein the peripheral nerve is disrupted, optionally by treatment with a protease (optionally collagenase or matrix metalloprotease) prior to culturing the peripheral nerve or cells from the peripheral nerve.

13 . The method of claim 7 , wherein the peripheral nerve is a sural nerve, a branch of a sural nerve, a proper digital nerve of a finger or toe, a gracilis branch of an obturator nerve, a segment of a medial antebrachial cutaneous nerve, a lateral antebrachial cutaneous nerve, a proximal third webspace fascicle nerve, or a posterior intraosseous nerve, optionally wherein the nerve is from an amputated limb.

14 . A method for producing a population of differentiated cells, comprising exposing a population according to claim 5 to differentiation conditions.

15 . The method of claim 14 , wherein:
(a) the differentiated cells are mesoderm cells and the differentiation conditions comprise culturing the population in a mesoderm differentiation medium; (b) the differentiated cells are endoderm cells and the differentiation conditions comprise culturing the population in an endoderm differentiation medium; or (c) the differentiated cells are ectoderm cells and the differentiation conditions comprise culturing the population in an ectoderm differentiation medium.

16 . The method of claim 14 , wherein:
(a) the differentiated cells are osteoblasts and the differentiation conditions comprise culturing the population in an osteogenic differentiation medium; (b) the differentiated cells are endothelial cells and the differentiation conditions comprise culturing the population in an endothelial differentiation medium; or (c) the differentiated cells are neural stem cells and the differentiation conditions comprise culturing the population in a neural stem cell differentiation medium.

17 . A population of differentiated cells produced by the method of claim 14 .

18 . A method of treating a subject in need of tissue regeneration, comprising implanting into the subject a population of cells suitable for said tissue regeneration, wherein the population of cells is a population of cells according to claim 5 , optionally wherein the population of cells is autologous to the subject.

19 . A method of treating a subject in need of gene therapy, comprising implanting into the subject a population of cells made recombinant by introduction of a gene of which the subject is in need, wherein the population of cells is a population of cells according to claim 5 , optionally wherein the population of cells is autologous to the subject.

Assignees

• Univ Kansas

Inventors

• Heggeness Michael H

Classifications

• C12N5/0607

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

• C12N2501/155

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

• C12N13/00 Primary

  Treatment of microorganisms or enzymes with electrical or wave energy, e.g. magnetism, sonic waves · CPC title

• C12N5/0623 Primary

  Stem cells · CPC title

• C12N2506/08

  from cells of the nervous system · CPC title