The Culture And Tracing Study In Vivo Of Rabbit Bone Marrow Mesenchymal Stem Cell

Objective:Using the adherent method to culture rabbit bone marrow mesenchymal stem cells (Bone Mesenchymal Stem Cells, BMSCs) in vitro. With the PLL@Fe3O4 to label the rabbit bone marrow mesenchymal stem cells (BMSCs) in vitro and inject the tagged cells through rabbit ear vein, taking MRI to understand the distribution of the cells in the liver. Providing the basic research results of MRI tracer for stem cell transplantation treatment.Methods:1. The rabbit BMSCs were cultured by adherent method. To confirm the obtained cells were BMSCs by the morphological observation and flow cytometry.2. To mark the BMSCs by PLL@Fe3O4.3. Prussian blue staining the transfected stem cells and observation of the situation of PLL@Fe3O4 markers in BMSCs.4. The effect of PLL@FesO4 on BMSCs was evaluated by trypan blue exclusion proliferation and MTT toxicity test.5. Taking PLL@Fe3O4 labeled rabbit BMSCs inject to the ear vein of rabbit and observing its distribution in the liver by MRI.Results 1. A large number of spindle and swirl adherent cells were observed by Inverted Phase Contrast Microscope. The high cells expressing of surface markers of marrow mesenchymal stem cells of CD44 and low cells expressing of surface markers of hematopoietic cells of CD 14 and CD45 were identified by flow cytometer, which confirmed the cultured cells were rabbit BMSCs.2. The rabbit BMSCs labeled by 20 μg/ml PLL@Fe3O4 were not found obvious damage and toxicity and PLL@Fe3O4 can be safely used in cell labeling.3. The PLL@Fe3O4 labeled rabbit BMSCs were found obviously to deposit in liver through the T2*W1 sequence detection of MRI, which lay the foundation of Stem cell tracking in vivo study. Meanwhile it is to prove that the bone marrow mesenchymal stem cells can go into the liver via peripheral vessels.