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 for producing a cellular composition comprising multipotent cells that express CD45 and HLA-ABC and are capable of differentiating into different types of non-hematopoietic cells 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 said multipotent cells.   
   
   
       2 . A method as claimed in  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 as claimed in  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 as claimed in  claim 1  which further comprises inducing the multipotent cells to differentiate into cells and tissues of non-hematopoietic lineages in vitro or in vivo. 
   
   
       5 . A cellular composition produced by a method of  claim 1 . 
   
   
       6 . A method of  claim 1  wherein positive selection is used to obtain an enriched hematopoietic CD45 + HLA-ABC +  cell preparation. 
   
   
       7 . A method of  claim 1  wherein negative selection is used to obtain an enriched hematopoietic CD45 + HLA-ABC +  cell preparation. 
   
   
       8 . 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. 
   
   
       9 . 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 pancreatic cells. 
   
   
       10 . A method of  claim 9  wherein the pancreatic cells are selected from the group consisting of acinar, ductal, islet-α, islet-β, islet-δ, and islet-PP cells. 
   
   
       11 . 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 muscle cells. 
   
   
       12 . An isolated and purified cellular composition comprising cells characterized by the following:
 (a) CD45 + HLA-ABC + ;   (b) round shape and non-adherent growth requirements; and   (c) stem cell factor receptor (KIT)+.   
   
   
       13 . An isolated and purified cellular composition of  claim 12  wherein the cells are further characterized by the following:
 (a) CD45 + HLA-ABC + ;   (b) Capable of differentiating into hematopoietic cells or progenitor cells;   (c) Capable of differentiating into one or more, two or more, three or more, four or more, five or more, or six or more, different non-hematopoietic cell types;   (d) round shape and non-adherent growth requirements;   (e) Stem cell factor receptor (KIT)+   (f) FLT3ligand receptor+;   (g) FGF receptor+;   (h) express embryonic stem cell proteins;   (i) HoxB4 + ;   (j) Flk-1 + ;   (k) CD34 ± ;   (l) non-tumorigenic;   (m) CD38 ± ; and   (n) derived from umbilical cord blood.   
   
   
       14 . A method of identifying the presence of cells of a cellular composition as claimed in  claim 12  in a mixed cell population comprising: exposing the cell population to an antibody or fragment thereof immunogenetically specific for markers (a) or (c) in  claim 12 , the occurrence of the markers being indicative of the presence of the cells in the cell population. 
   
   
       15 . A method for obtaining non-hematopoietic cells for autologous transplantation from a patient's own hematopoietic cells comprising (a) obtaining a sample comprising hematopoietic cells from the patient; (b) separating out an enriched cell preparation comprising CD45 + HLA-ABC +  cells; (b) culturing the cells in a medium comprising FGF-4, SCF and FLT-3 ligand to produce multipotent cells with the potential or increased potential to form non-hematopoietic cells. 
   
   
       16 . An isolated and purified cellular composition comprising multipotent cells that express CD45, HLA-ABC and OCT-4 and are capable of differentiating into different types of non-hematopoietic cells. 
   
   
       17 . A pharmaceutical composition comprising a cellular composition of  claim 16 , and a pharmaceutically acceptable carrier, excipient, or diluent. 
   
   
       18 . A method of treating a patient with a condition involving non-hematopoietic cells comprising transferring a cellular composition of  claim 16  into the patient, wherein the cells in the composition differentiate into the non-hematopoietic cells. 
   
   
       19 . A method of  claim 18  wherein the non-hematopoietic cells are pancreatic cells and the condition is diabetes. 
   
   
       20 . An isolated and purified cellular composition comprising multipotent cells that express CD45, HLA-ABC, OCT-4, GFAP, desmin, nestin and Flk-1 and are capable of differentiating into different types of non-hematopoietic cells.