Identification of cells at early and late stages of polarization during odontoblast differentiation using pOBCol3.6GFP and pOBCol2.3GFP transgenic mice

• Published in: Bone (New York, N.Y.) N.Y.), 2010-11, Vol.47 (5), p.948-958
• Main Authors: Balic, Anamaria, Aguila, H. Leonardo, Mina, Mina
• Format: Article
• Language: English
• Subjects: Animal models; Animals; Cell Cycle - genetics; Cell Cycle - physiology; Cell Differentiation - physiology; Cells, Cultured; Collagen Type I - metabolism; Dental Pulp - cytology; Flow Cytometry; Green Fluorescent Proteins - genetics; Green Fluorescent Proteins - metabolism; Immunohistochemistry; Mice; Mice, Transgenic; Microscopy, Confocal; Odontoblasts - cytology; Odontoblasts - metabolism; Orthopedics
• ISSN: 8756-3282; 1873-2763; 1873-2763
• DOI: 10.1016/j.bone.2010.08.009

Abstract Transgenic mouse lines in which GFP expression is under the control of tissue- and stage specific promoters have provided powerful experimental tools for identification and isolation of cells at specific stage of differentiation along a lineage. In the present study, we used primary cell cultures derived from the dental pulp from pOBCol3.6GFP and pOBCol2.3GFP transgenic mice as a model to develop markers for early stages of odontoblast differentiation from progenitor cells. We analyzed the temporal and spatial expression of 2.3-GFP and 3.6-GFP during in vitro mineralization. Using FACS to separate cells based on GFP expression, we obtained relatively homogenous subpopulations of cells and analyzed their dentinogenic potentials and their progression into odontoblasts. Our observations showed that these transgenes were activated before the onset of matrix deposition and in cells at different stages of polarization. The 3.6-GFP transgene was activated in cells in early stages of polarization, whereas the 2.3-GFP transgene was activated at a later stage of polarization just before or at the time of formation of secretory odontoblast.