Culturing And Evaluating Of The SDF-1 Modified Human Bone Marrow Mesenchymal Stem Cells In Vitro

Objective:The injury of Hematopoietic microenvironment (HME) is the important reason for hematologic diseases and the delay of rebuilding hematopoiesis during the hematopoietic stem cells transplantation. Marrow stromal cells,the mostly components of the HME , can adhere the hematopoietic stem/progenitor cells and enhance their homing,proliferation and differentiation. Bone mesenchymal stem cells (BMSCs), which are different from hematopoietic stem cells (HSCs), are another kind of bone marrow stem cells. BMSCs can not only support the hemopoiesis by participating in the formation of hemopoietic micro-environment, but also differentiate to different marrow stromal cells which can also support hematopoiesis in the suitable conditions. BMSCs are the perfect cells'source of cellular transplant and gene treatments for the characteristics of self-renewal and multi-directional differentiating abilities. They can recover and improve the injured HIM and promote to rebuild hematopoiesis and immunity during the hematopoietic stem cells transplantation.Stromal cell derived factor-1 (SDF-1), a significant component in hematopoietic microenvironment, is mostly synthesized and secreted by bone marrow stromal cells. Human SDF-1, whose gene locates at chromosome ten, and classified as a member of chemokine CXC subfamily according to its amino acid sequence, takes part in the regulation and control of hematopoiesis through binding to its special receptor CXCR4 on the surface of hematopoietic cells. SDF-1 not only plays important roles in the mobilization, homing, location and proliferation of hematopoietic stem cells, but also can maintain the survival of hematopoietic stem cells depend itself or through cellular signal transduction system by cooperating with other cytokines such as G-CSF,TPO .In our experiment, we successfully isolated and cultured the human BMSCs (hBMSCs) and then evaluated them, after that, we constructed the eukaryotic expression vector of SDF-1 and then transfected it to hBMSCs and tested the instantaneous transferred efficiency. All our experiment is the prior basic study for the following in vitro and in vivo...