Isolation, culture and identification of bovine cementum-derived cells / 中华口腔医学杂志

ABSTRACT

<p><b>OBJECTIVE</b>To isolate and culture bovine cementum-derived cells and investigate their biological properties.</p><p><b>METHODS</b>Culture of primary bovine cementoblast (CB) were established from new-born bovine teeth. Cementum was manually dissected, fragmented, and digested twice with collagenase. Following a thorough wash to remove liberated cells, the remaining cementum fragments were plated and cultured in RPMI1640 containing 10% FCS and 1.5% Hepes. The cells in culture were identified using immunocytochemistry by expression of cementum attachment protein (CAP), osteocalcin (OCN) and alkaline phosphatase (ALP). ALP activity was measured by modified Gomori staining.</p><p><b>RESULTS</b>(1) The cells in culture possessed the typical morphology of CB and there were no obvious change of the cell morphology up to 5th passage; (2) Cells in culture exhibited alkaline phosphatase activity and were also positive to CAP, OPN and ALP immunohistochemistry staining.</p><p><b>CONCLUSION</b>The cells isolated and cultured in this experiment possess the properties of CB. This enables us to further observe its biological characteristics during cementogenesis and periodontal regeneration.</p>