The effects of OPG and RANKL expressed on HDFCs

Objective:To establish the co-culture system of human dental follicle cells (HDFCs) and human peripheral blood mononuclear cells (PBMCs) ,and study the effects of OPG and RANKL expressed on human dental follicle cells on osteoclastogenesis. Methods: Mononuclear cells ,separated from healthy human peripheral blood, were seeded on 24-well plates and cultured in 1640 medium.A stainless steel net was placed in each well in advance,and HDFCs growing on a cover slip were placed on the net. So in this system the two type of cells did not contact,while the moleculars can communicate in the medium.The study includes 7 groups:AAPBMCs; BAPBMCs+ HDFCs (contacted); CAPBMCs+ HDFCs (contacted) +CSF-1+PTHrP; DAPBMCs+ HDFCs (uncontacted); EAPBMCs+ CSF-1+RANKL; FAPBMCs+ CSF-1+RANKL+ HDFCs (uncontacted) ; GAPBMCs+ CSF-1+RANKL+ HDFCs (uncontacted) {anti-OPG..The cells of each group were seeded on the coverslips and dentin slices,placed in the wells in advance,and cultured in 1640 medium.At 10 days the PBMCs, growing on coverslips, were treated for TRAP staining; At 14 days the dentin slices were taken out and the resorption pits were observed by scaning electron microscope. Results:There are few osteoclast like cells(OLCs) in group B and F,and no OLCs were found in group A and C. The OLCs in group E are most ,followed by group C and group G..Conclusions: We establish the co-culture system of two types of cells and prove the effects of OPG and RANKL expressed on human dental follicle cells on osteoclastogenesis.