Primary Culture and Identification of Osteoblasts from Subchondral Bone of Neonatal SD Rat's Condylar Process / 昆明医科大学学报

ABSTRACT

Objective Establish an experimental model for primary subchondral bone osteoblasts culture of neonatal SD rat's condylar process. Methods Under the condition of sterile,24-hour SD rat was executed and its condylar process was isolated. Removing cartilage layer, the subchondral bone was exposed obviously, then it was cultured with modified repeating enzymatic digestion-adherent explants method. The cellular morphology was identified with invert microscope and immunohistochemistry staining, the osteoblasts were identified by alkaline phosphorase (ALP) staining and calcified nodules staining, and the proliferation of the acquired cells was examined by methyl thiazolyl tetrazolium (MTT) assay. Results A variety of cell morphologies were observed, such as spindle-shaped, triangular and irregular-shape, and their cell processes were significant. The alkaline phosphatase staining and calcified nodules staining of cultured osteoblasts with mineralized nodules were positive. Cells grew slowly in 1-3 days, and the cells growth reached the highest level at the 8th day. The cells growth trend has gradually slowed down after 8 days. Conclusion The method is an efficient way to culture and obtain purified neonatal SD rat's subchondral bone osteoblasts with typical characteristics.