Changes Of Osteoblasts And Osteoclasts During G-CSF Induced Mobilization Of Hematopoietic Stem Cells In Mice

Objective: hematopoietic stem cells(Hematopoietic stem cell HSC)has been widely used for transplantation in the treatment of malignant hematological diseases,collecting a sufficient amount of hematopoietic stem cell transplantation is key prerequisite for the success of recent studies clearly,there are more than two kinds of bone marrow niches in bone marrow microenvironment,the endosteal niche to maintain hematopoietic stability a cell plays an important role in the osteoblast is an important part of the endosteal niche,a large number of experiments show that G-CSF mobilization after reducing the number of osteoblast,osteoclast function decreased,as the reaction with the osteoblasts cells,what happens in the mobilization process,is worth us to study.In this study,mice as the research object,observe the mobilization of G-CSF osteoblasts and osteoclasts of mice,and the activation of osteoclasts induced by PTH,to observe the fine stem Effects of cell mobilization.method:Using flow cytometry in 0,3,5 days of mobilization of peripheral blood stem cells in mice were induced by G-CSF proportion,mobilization,throughreal-time quantitative polymerase chain reaction(RQ-PCR)mobilization of bone marrow cells before and after detection of osteocalcin(Osteocalcin OCN),stromal cell derived factor-1(Stromal cell-derived factor-1 SDF-1)and matrix metalloproteinase 9(Matrix metalloproteinases 9 MMP9)the expression of mRNA,immunohistochemical detection of bone marrow OCN 0 Chemistry(osteoblast specific expression)the number of positive cells to compare before and after mobilization into changes in the number and function of bone cells of mice peripheral blood TRACP-5b concentration was detected by ELISA osteoclast activity,tartrate resistant acid phosphatase(tartrate-resistant acid phosphatase TRAP)the number of osteoclast was detected in mouse femur,evaluation of G-CSF mobilization on bone,broken Effects of bone cells.Large dose of PTH(hormone)of parathyroid hormone(Parathyroid)can activate osteoclasts to observe whether HSC activation can be induced by activated osteoclasts.Result:1.Bone marrow SDF-1 and OCN expression decreased,MMP9 expression increased 0 days(0.9 + 0.21),3 days(0.72 + 0.62),5 days(2.13 + 0.71)suggested that osteoblast function decreased,neutrophils release MMP9,bone marrow stromal protein hydrolysis.2.Under microscope,the number of bone marrow cells was decreased and the morphology changed from the original ovoid to fusiform to flat.3.TRACP-5b increased in peripheral blood of mice,and was increased at day 0(+ +.0.52IU/L)for 5 days(+ 10.49 + 1.75mIU/mL),P<0.05.TRAP staining showed that the number of osteoclasts increased and the activity increased after G-CSF injection.4.Doses of PTH activated osteoclasts can produce HSC mobilizationeffect.Conclusion:After G-CSF mobilization,the number and activity of osteoblasts decreased,resulting in SDF-1 chemotactic ability is abate,destruction of hematopoietic stem cell quiescence induced mobilization;osteoclast activation after interaction with osteoblasts influence mobilization occurred.