Amplication and identification of dendritic cells from rat bone marrow in vitro

To establish the method of inducing and proliferating dentritic cells (DCs) from rat bone marrow in vitro and analyze its biological characteristics. Bone marrow cells were flushed from femurs and tibias of healthy Lewis rats, depleted of erythrocytes with osmotic lysis. Cultured for 48 hours, the floating cells were removed and the adherent cells were continued to culture with cytokines including IL-4 and GM-CSF. After 14 days culture, the induced DCs were observed by photics microscope and electron microscope, also co-cultured with native T cells derived from spleen, its stimulating function was detected by MTT assay. The induced DCs had the typical morphology characteristitics. Only a little number of the induced DCs can stimulate the proliferation of allogeneic T cells efficiently and they were potent stimulators in allogeneic mixed lymphocyte reaction (MLR). [Conclusion] It is an applicable method by using rat bone marrow as a source after adherent and being cultured in the medium containing IL-4 and GM-CSF, which could generate plenty of DCs with high purity and biological functions.