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[Isolation, culture and identification of bovine cementum-derived cells]
Jianxia Hou1, Caifang Cao, Huanxin Meng
1Department of Periodontology, Peking University School of Stomatology, Beijing 100081, China.
Objective:
To isolate and culture bovine cementum-derived cells and investigate their biological properties.
Methods:
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.
Results:
(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.
Conclusion:
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.