Cellular Compositions and Methods of Making and Using Them

Abstract

The invention relates to cellular compositions comprising hematopoietic cells with the potential or increased potential to form non-hematopoietic cells; methods for producing such cellular compositions; methods for differentiation of cells of cellular compositions of the invention into cells that exhibit morphological, physiological, functional, and/or immunological features of non-hematopoietic cells; and uses of the cellular compositions. The invention also relates to a method for the expansion of hematopoietic stem and progenitor cells.

1 . A method comprising:
 (a) enriching hematopoietic cells to obtain an enriched hematopoietic CD45 + HLA-ABC +  cell preparation;   (b) culturing the enriched hematopoietic CD45 + HLA-ABC +  cell preparation in a medium comprising FGF-4, SCF and FLT-3 ligand to provide a cellular composition comprising multipotent cells that express CD45 and HLA-ABC.   
     
     
         2 . A method of  claim 1  wherein the hematopoietic cells are obtained from a source selected from the group consisting of blood, blood fractions, bone marrow and umbilical cord blood. 
     
     
         3 . A method of  claim 1  wherein the enriched hematopoietic cell preparation is enriched in CD45 + HLA-ABC + CD2 − CD3 − CD14 − CD16 − CD19 − CD24 − CD56 − CD66 −  glycophorin A −  cells. 
     
     
         4 . A method of  claim 1  further comprising transferring the composition comprising multipotent cells into a patient. 
     
     
         5 . A method of  claim 4  wherein the hematopoietic cells in (a) are obtained from a sample from the patient. 
     
     
         6 . A method of  claim 4  wherein the hematopoietic cells in (a) are obtained from a sample from an individual other than the patient. 
     
     
         7 . A method of  claim 1  further comprising culturing the multipotent cells in the presence of a differentiation factor that induces differentiation of the multipotent cells into cells expressing glucagon, insulin, somatostatin and/or pancreatic polypeptide. 
     
     
         8 . A cellular composition produced by a method of  claim 1 . 
     
     
         9 . An isolated and purified cellular composition comprising:
 (a) cells characterized by the following: CD45 + HLA-ABC + , round shape and non-adherent growth requirements, and stem cell factor receptor (KIT) + ; or   (b) multipotent cells that express CD45 and HLA-ABC and are capable of differentiating into different types of non-hematopoietic cells.   
     
     
         10 . An isolated and purified cellular composition of  claim 9  wherein the cells are further characterized by the following: (c) capable of differentiating into hematopoietic cells or progenitor cells; (d) capable of differentiating into different non-hematopoietic cell types; (e) FLT3ligand receptor+; (f) FGF receptor+; (g) express embryonic stem cell proteins; (h) HoxB4 + ; (i) Flk-1 + ; (k) CD34 ± ; (j) non-tumorigenic; (k) CD38 ± ; and (l) derived from umbilical cord blood. 
     
     
         11 . A method of identifying the presence of cells of a cellular composition as claimed in  claim 9  in a mixed cell population comprising: exposing the cell population to an antibody or fragment thereof immunogenetically specific for markers CD45 + , HLA-ABC + , or KIT + , the occurrence of the markers being indicative of the presence of the cells in the cell population. 
     
     
         12 . An isolated cellular composition comprising multipotent cells that express CD45 and HLA-ABC and are capable of differentiating into different types of non-hematopoietic cells. 
     
     
         13 . An isolated cellular composition of  claim 12  wherein the non-hematopoietic cells are pancreatic cells. 
     
     
         14 . An isolated cellular composition of  claim 12  wherein the multipotent cells express OCT-4, desmin, nestin or Flk-1. 
     
     
         15 . An isolated cellular composition of  claim 12  wherein the multipotent cells express OCT-4, GFAP, desmin, nestin and Flk-1. 
     
     
         16 . A pharmaceutical composition comprising a cellular composition of  claim 12 , and a pharmaceutically acceptable carrier, excipient, or diluent.