Soluble factor cross-talk between human bone marrow-derived hematopoietic and mesenchymal cells enhances in vitro CFU-F and CFU-O growth and reveals heterogeneity in the mesenchymal progenitor cell compartment.
The homeostatic adult bone marrow (BM) is a complex tissue wherein physical and biochemical interactions serve to maintain a balance between the hematopoietic and nonhematopoietic compartments. To focus on soluble factor interactions occurring between mesenchymal and hematopoietic cells, a serum-free adhesion-independent culture system was developed that allows manipulation of the growth of both mesenchymal and hematopoietic human BM-derived progenitors and the balance between these compartments. Factorial experiments demonstrated a role for stem cell factor (SCF) and interleukin 3 (IL-3) in the concomitant growth of hematopoietic (CD45+) and nonhematopoietic (CD45-) cells, as well as their derivatives. Kinetic tracking of IL-3alpha receptor (CD123) and SCF receptor (CD117) expression on a sorted CD45- cell population revealed the emergence of CD45-CD123+ cells capable of osteogenesis. Of the total fibroblast colony-forming units (CFU-Fs) and osteoblast colony-forming units (CFU-O), approximately 24% of CFU-Fs and about 22% of CFU-Os were recovered from this population. Cell-sorting experiments demonstrated that the CD45+ cell population secreted soluble factors that positively affect the survival and proliferation of CFU-Fs and CFU-Os generated from the CD45- cells. Together, our results provide insight into the intercellular cytokine network between hematopoietic and mesenchymal cells and provide a strategy to mutually culture both mesenchymal and hematopoietic cells in a defined scalable bioprocess.
Baksh
Dolores
D
Institute of Biomaterials and Biomedical Engineering, Department of Chemical Engineering and Applied Chemistry, Faculty of Dentistry, University of Toronto, Rm 407, Roseburgh Bldg, 4 Taddle Creek Rd, Toronto, ON, M5S 3G9, Canada.
Davies
John E
JE
Zandstra
Peter W
PW
D002454:Cell Differentiation; D019070:Cell Lineage; D049109:Cell Proliferation / Q000187:drug effects; D002478:Cells, Cultured; D003114:Colony-Forming Units Assay; D016895:Culture Media, Serum-Free; D005347:Fibroblasts; D006412:Hematopoietic Stem Cells / Q000166:cytology* / Q000378:metabolism; D006801:Humans; D007377:Interleukin-3 / Q000494:pharmacology; D053650:Interleukin-3 Receptor alpha Subunit; D017493:Leukocyte Common Antigens / Q000378:metabolism; D059630:Mesenchymal Stem Cells / Q000166:cytology* / Q000378:metabolism; D010006:Osteoblasts / Q000166:cytology*; D016185:Receptors, Interleukin-3 / Q000378:metabolism; D019089:Stem Cell Factor / Q000494:pharmacology
D016895:Culture Media, Serum-Free; C115022:IL3RA protein, human; D007377:Interleukin-3; D053650:Interleukin-3 Receptor alpha Subunit; D016185:Receptors, Interleukin-3; D019089:Stem Cell Factor; D017493:Leukocyte Common Antigens