[Culture and identification of dendritic cells derived from bone marrow of DA rats]

Objective To isolate, culture and identify bone marrow-derived dendritic cells (BMDCs) in vitro from DA rats. Methods Bone marrow cells were removed from the bone marrow cavity between femur and tibia in DA rats. The cells were induced with 20 ng/mL recombinant mouse granulocyte macrophage colony stimulating factor (rmGM-CSF) and 10 ng/mL recombinant mouse interleukin 4 (rmIL-4) in vitro for 7 days for differentiation into DCs. The level of IL-12 in the culture supernatant of immature dendritic cells (imDCs) was detected by ELISA. The proliferation and morphological changes of BMDCs were observed under an inverted microscope, the morphology of DCs was observed by scanning electron microscope. The expression of surface specific markers OX62/CD103, CD40, CD80, CD86 and MHC II in imDCs was analyzed by flow cytometry. Mixed lymphocyte reaction (MLR) was used to detect the proliferation ability of T lymphocytes induced by imDCs. Results A large amount of imDCs were obtained from DA rat bone marrow cells cultured in vitro. The expression of OX62 was high and the expression of CD40, CD80, CD86 and MHC- II were low in the imDCs. The content of IL-12 in the supernatant of imDCs treated with rmIL-10 decreased with the extension of culture time, but it was the opposite in the group treated with rmIL-4, and the ability of imDCs to induce T lymphocyte proliferation was weak. Conclusion A simple method for the isolation and culture of high-purity imDCs from DA rats has been successfully established.