Materials and methods for generating therapeutic mesenchymal stem cells

Abstract

Embodiments of the present disclosure relate generally to the production of therapeutic mesenchymal stem cells (MSCs). More particularly, the present disclosure relates to the use of cell culture compositions and methods for generating MSCs that secrete neurotrophic factors and synaptic organizing agents for the treatment of neurodegenerative diseases such as Amyotrophic Lateral Sclerosis (ALS). As such, the present disclosure addresses the need for establishing a reliable source of therapeutic stem cells useful for the treatment of neurodegenerative diseases.

What is claimed is: 
     
         1 . An isolated non-genetically modified human cell activated ex vivo from a mesenchymal stem cell (MSC) under conditions such that the isolated non-genetically modified human cell secretes laminin β2 at a level that is greater than the basal secretion level of laminin β2 by the MSC. 
     
     
         2 . The isolated human cell of  claim 1 , wherein the MSC is obtained from Wharton's jelly from an umbilical cord. 
     
     
         3 . The isolated human cell of  claim 1 , wherein the MSC is CD140b positive. 
     
     
         4 . The isolated human cell of  claim 1 , wherein the cell is activated in a cell culture composition comprising cell culture media and Insulin-like growth factor 1 (IGF-1) 
     
     
         5 . The isolated human cell of  claim 4 , wherein the cell culture composition further comprises one or more additional growth factors selected from Fibroblast Growth Factor (FGF) and Platelet-derived Growth Factor (PDGF). 
     
     
         6 . The isolated human cell of  claim 4 , wherein the cell culture composition further comprises Heregulin β1. 
     
     
         7 . The isolated human cell of  claim 4 , wherein the cell culture media is DMEM-F12 containing L-glutamine. 
     
     
         8 . The isolated human cell of  claim 4 , wherein the cell culture composition further comprises dibutryl cAMP. 
     
     
         9 . The isolated human cell of  claim 4 , wherein the cell culture composition further comprises 3-isobutyl-1-methylxanthine (IBMX). 
     
     
         10 . The isolated human cell of  claim 1 , wherein the isolated non-genetically modified human cell further secretes one or more of glial cell-derived neurotrophic factor (GDNF), brain-derived neurotrophic factor (BDNF), and vascular endothelial growth factor (VEGF) at a level that is greater than the basal secretion level of GDNF, BDNF, or VEGF by the MSC. 
     
     
         11 . The isolated human cell of  claim 1 , wherein the isolated human cell enhances growth and/or survival of one or more motor nerve terminals at a neuromuscular junction upon exposure to the activated MSCs. 
     
     
         12 . The isolated human cell of  claim 1 , wherein the isolated human cell ameliorates denervation at a neuromuscular junction caused by Amyotrophic Lateral Sclerosis (ALS). 
     
     
         13 . A method of treating a disease for which administration of neurotrophic factors is beneficial in a subject in need thereof, comprising administering to the subject the isolated human cell of  claim 1 . 
     
     
         14 . The method of  claim 13 , wherein the diseases is ALS. 
     
     
         15 . A method for producing a cell from a mesenchymal stem cell (MSC), such that the cell secretes laminin β2 at a level that is greater than the basal secretion level of laminin β2 by the MSC, the method comprising:
 exposing the MSC to a cell culture composition comprising cell culture media and Insulin-like growth factor 1 (IGF-1). 
 
     
     
         16 . The method of  claim 15 , wherein the MSC is obtained from Wharton's jelly from an umbilical cord. 
     
     
         17 . The method of  claim 15 , wherein the MSC is CD140b positive. 
     
     
         18 . The method of  claim 15 , wherein the cell culture composition further comprises one or more additional growth factors selected from Fibroblast Growth Factor (FGF) and Platelet-derived Growth Factor (PDGF). 
     
     
         19 . The method of  claim 15 , wherein the cell culture composition further comprises Heregulin β1. 
     
     
         20 . The method of  claim 15 , wherein the cell culture media is DMEM-F12 containing L-glutamine. 
     
     
         21 . The method of  claim 15 , wherein the cell culture composition further comprises dibutryl cAMP. 
     
     
         22 . The method of  claim 15 , wherein the cell culture composition further comprises 3-isobutyl-1-methylxanthine (IBMX). 
     
     
         23 . The method of  claim 15 , wherein the cell further secretes one or more of glial cell-derived neurotrophic factor (GDNF), brain-derived neurotrophic factor (BDNF), and vascular endothelial growth factor (VEGF) at a level that is greater than the basal secretion level of GDNF, BDNF, or VEGF by the MSC. 
     
     
         24 . The method of  claim 15 , wherein the cell enhances growth and/or survival of one or more motor nerve terminals at a neuromuscular junction upon exposure to the cell. 
     
     
         25 . The method of  claim 15 , wherein the cell ameliorates denervation at a neuromuscular junction caused by Amyotrophic Lateral Sclerosis (ALS).