Aim: To induce and expand dendritic cells (DC) from rat bone marrow in vitro and identify their biological characterization.
Methods: The rat bone marrow cells were collected and cultured for 48 hours and the floating cells were removed. Then IL-4 and GM-CSF were added into the fresh medium. After 2 weeks, the morphological character of the cultured DCs was observed under light microscope and transmission electron microscope. Expressions of MHC class II molecule, B7-1 and B7-2 were detected by flow cytometry. The cultured DCs were co-cultured with allogenic T cells derived from rat spleen. T cell proliferation was measured by MTT colorimetry.
Results: The cultured DCs had the typical morphological characterization of DC, and the expression rates of MHC class II molecule, B7-1 and B7-2 were 74.2%, 81% and 76% respectively. The cultured DCs could notably stimulate the proliferation of allogeneic T cells.
Conclusion: The adherent culture of rat bone marrow cells, and co-culture with IL-4 and GM-CSF can obtain a number of high purity of DCs, which lay the foundation for study on DC's function.