Method for purifying pluri-differentiated mesenchymal progenitor cells
There is provided an isolated pluri-differentiated human mesenchymal progenitor cells (MPCs), a method for isolating and purifying human mesenchymal progenitor cells from Dexter-type cultures, and characterization of and uses, particularly therapeutic uses for such cells. Specifically, there is provided isolated MPCs which can be used for diagnostic purposes, to enhance the engraftment of hematopoietic progenitor cells, enhance bone marrow transplantation, or aid in the treatment or prevention of graft versus host disease.
1. A method for purifying pluri-differentiated mesenchymal progenitor cells comprising the steps of:
a) providing a cell culture preparation by the Dexter method;
b) treating the cells to obtain a cell suspension;
c) removing macrophages;
d) fractionating the cells; and
e) collecting the fraction of pluri-differentiated mesenchymal progenitor cells.
2. The method of claim 1 , wherein said fractionating of step d) is carried out by discontinuous Percoll gradient centrifugation.
3. The method of claim 1 , wherein the collected fraction of pluri-differentiated mesenchymal progenitor cells has a purity of at least 95%.
4. The method of claim 1 , wherein the pluri-differentiated mesenchymal progenitor cells are human cells.
5. The method of claim 1 , wherein the pluri-differentiated mesenchymal progenitor cells each simultaneously expresses, at the protein level, a plurality of genes that are markers for multiple cell lineages, wherein the multiple cell lineages comprise at least four different mesenchymal cell lineages, and wherein each of the markers is specific for a single cell lineage.
6. The method of claim 5 , wherein the at least four different mesenchymal cell lineages comprise adipocyte, osteoblast, fibroblast, and muscle cell.
7. The method of claim 5 , wherein the markers specific for a single cell lineage are selected from the group consisting of Nile Red, Oil Red O, adipsin, alkaline phosphatase, cadherin-11, chondroitin sulfate, collagen type I, decorin, fibronectin, prolyl-4-hydroxylase, actin, caldesmon, and transgelin.
8. The method of claim 1 , wherein the pluri-differentiated mesenchymal progenitor cells are not neoplastic cells.
9. The method of claim 1 , wherein the pluri-differentiated mesenchymal progenitor cells are chromosomally normal, as determined by Geimsa-trypsin-Wrights (GTW) banding.