Biological Characterization Of Bone Marrow Derived Mesenchymal Stem Cells From Leukemia Patients

Objective To explore the biological characteristics of bone marrow derived mesenchymal stem cells (MSC) from leukemia patients, and examine their multiple differentiation capacity, immunophenotype and expression of leukemia-related oncogene in order to provide experimental basis for clinical application of cell based therapy.Methods MSC were obtained from leukemia patients and healthy adults bone marrow and cultured in expanded medium. The cell cycle and immunophenotype of the expanded MSC were detected by fluorescence-activated cell sorter (FACS). The growth curve was investigated. Bone marrow drived MSC from leukemia patients and normal adults can be differentiated into osteocyte and adipocyte by different agents, which were identified by Von Kossa stain and Oil-red O staining. RT-PCR was used to detect the expression of leukemia-related oncogene and the gene of hemopoietic growth factor such as IL-6, IL-3, SCF and SDF-1. Content of IL-6 and TNF-a in supernatant of cultivation were detected by radio-immunization. MSC were culture in semisolid culture containing hemopoietic growth factor for 4 to 5 weeks to observe the hematopoietic cell colony formation.Results MSC from leukemia patients and normal adults were similar in cell morphology, growth pattern and immunophenotype. They all showed a typical fibroblast like morphology. The cultured MSC expressed CD29, CD44, CD73 and CD 105, but not CD31, CD34, CD45 and HLA-DR. After 4 to 5 weeks'culture, no hematopoietic cell colony formation was observed from the MSC. FACS revealed that more than 90% MSC derived from leukemia patients were at G₀/G₁. Under suitable conditions, Leukemia patients derived MSC could differentiate into adipocyte and osteoblast. Moreover, MSC derived from leukemia patients showed a normal the gene expression of hemopoietic growth factor, and secretion of IL-6 and TNF-a were...