Experimental Study Of TPO Gene Transfected Into Bone Marrow Stromal Cells In Mice To Promote Hematopoietic Reconstitution

Objective: Hematopoietic stem cell transplantation (HSCT) is a very important method for curing hematologic malignancy and some solid carcinomas sensitive to radiotherapy/chemotherapy, as well as acute radiation diseases. Delays of hematopoietic reconstitution and hematopoietic depression after HSCT lead to decrease of white blood cell counts, platelet counts. Infection and bleeding are major complications posttransplantation, also they are primary causes associated with transplantation-related mortality. At present, it is confirmed the delay of hematopoietic reconstitution is associated with the injury of bone marrow stromal cells (BMSCs), deficient in hematopoietic microenvironment that can improve proliferation and differentiation of hematopoietic stem progenitor cell. Since the application of granulocyte colony-stimulating factor (G-CSF), granulocyte-macrophage colony-stimulating factor (GM-CSF) in clinic, it has enhanced the neutrophil recovery notably and cut down the incidence of infection. However, it depends mainly on infusing platelet suspension with respect to acute bleeding. While, a long-term infusing platelet suspension maybe bring to many severe side effects, such as costly expense. It also perhaps develops platelet antibody because of infusing platelet suspension again and again. It is confirmed that thrombopoietin (TPO) is a primary, special factor to regulate thrombopoiesis. So, we selected BMSCs as target cells, TPO cDNA eukaryotic expressing plasmid was transfected into BMSCs with Lipofectamine 2000. By observing the effects on improving platelet recovery and hematopoietic reconstitution by engrafting BMSCs transfected TPO gene, we explored the gene therapy feasibility of hematopoietic growth factor mediated by BMSCs and the effects on promoting hematopoietic reconstitution.Methods: In vitro, BMSCs of the mice were separated and cultured referring to Dexter-type long-term marrow cultures (D-LTMC). BMSCs cultured primarily were confirmed by immunocytochemistry and cytochemical staining metheds. TPO cDNA eukaryotic expressing plasmid was transfected into BMSCs with Lipofectamine 2000,...