Identification of cells at early and late stages of polarization during odontoblast differentiation using pOBCol3.6GFP and pOBCol2.3GFP transgenic mice |
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| Authors: | Anamaria Balic H. Leonardo Aguila Mina Mina |
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| Affiliation: | 1. Department of Craniofacial Sciences, School of Dental Medicine, University of Connecticut Health Center, Farmington, CT, USA;2. Department of Immunology, School of Medicine, University of Connecticut Health Center, Farmington, CT, USA;1. Department I of Anatomy, University of Cologne, Cologne, Germany;2. Department of Operative Dentistry, Periodontics and Endodontics, Heinrich-Heine-University, Düsseldorf, Germany;1. Hospital of Stomatology, Zunyi Medical College, Zunyi, Guizhou, People''s Republic of China;2. Department of Endodontics, Fourth Military Medical University, Xian, Shanxi, People''s Republic of China;3. Department of Stomatology, Xian Ninth Hospital, Xian, Shanxi, People''s Republic of China;4. Center of Oral Disease, 306th Hospital, Beijing, People''s Republic of China;1. The State Key Laboratory Breeding Base of Basic Science of Stomatology (Hubei-MOST) and Key Laboratory of Oral Biomedicine Ministry of Education, School and Hospital of Stomatology, Wuhan University, Wuhan, China;2. Department of Pediatric Dentistry, School and Hospital of Stomatology, Wuhan University, Wuhan, China;3. Department of Endodontics, University of California Los Angeles, Los Angeles, California;4. Department of Periodontology, School of Dental Medicine, University of Bern, Bern, Switzerland;6. Department of Dental Implantology, School and Hospital of Stomatology, Wuhan University, Wuhan, China;1. Department of Oral Histology-Developmental Biology, School of Dentistry and Dental Research Institute, BK 21, Seoul National University, Seoul 110-749, Republic of Korea;2. Department of Oral and Maxillofacial Surgery, School of Dentistry, Seoul National University, Seoul 110-749, Republic of Korea |
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| 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. |
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