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

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.

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.