The Establishment of Fibronectin-mediated Haemal Tumor Chemotherapy Resistance Model Simulating the Bone Marrow Microenviroment in Vivo
|School||Huazhong University of Science and Technology|
|Keywords||Blood tumor Bone marrow microenviroment Chemotherapy-resistance Fibronectin|
Objective:1. To Separate and identificate the bone marrow stromal cells.2. To detect whether fibronectin exist or is expressed in bone marrow stromal cells.3. To make the blood tumor cell contact with fibronectin simulating the bone marrow microenvironment in vivo, to validate if fibronectin has mediated the minimal residual disease of blood tumor cells, namely tumor resistance.Methods:1. To collect the bone marrow aspiration specimens of non-malignant tumor patients, by the method of density gradient centrifugalization to obtain bone marrow stromal cells and culture them which adhere on the cultural bottles’plastic surface . To observe the morphous of the BMSCs directly under the microscope and then to detect the symbols of the BMSCs by flow cytometric analysis.2. To detect the expression of fibronectin on or in BMSCs.3. To make the K562 cell-line co-cultured with the fibronectin wrapped plateand MSCs, respectively.Then to add imatinib or daunorubicin into those two co-cultural systems mentioned above.Finally to detect the cell cycle arrest and apoptosis. Results:1. We have separated and cultured MSCs , which expressed CD44,CD105 positive(PE marks), and CD19,CD45,CD34 negative(FITC marks).2. Fn has been detected in MSCs’cytoplasm,which can be secreted from them,either.3. Fn have induced K562 cell in G1 stage arrest and reduced apoptosis rate.Conclusions:1. We have separated and cultured MSCs ,which can be used to establish Fn-mediated adhesion resistance models. The molecular markers of these MSCs which can be used for further research are coincidence with international studies.2. Fn have been detected in MSCs’cytoplasm, and can be secreted from them either.3. MSCs might induce blood tumor chemotherapy adhesion resistance by secreting Fn.