Ju Eun Oh1, Jin-Kyu Yi2,3. 1. Anesthesia and Pain Research Institute, School of Medicine, Yonsei University, Seoul, South Korea. 2. Department of Conservative Dentistry, School of Dentistry, Kyung Hee University, Seoul, South Korea. jink.yi@khu.ac.kr. 3. Department of Conservative Dentistry, Kyung Hee University Dental Hospital at Gangdong, 892, Dongnam-Ro, Gangdong-Gu, Seoul, 05278, South Korea. jink.yi@khu.ac.kr.
Abstract
OBJECTIVES: The aim of this study was the isolation and characterization of dental follicle-derived Hertwig's epithelial root sheath cells (DF-HERSCs). MATERIALS AND METHODS: DF-HERSCs were isolated from dental follicle (DF)-derived single-cell suspensions. Their epithelial phenotypes were analyzed by Western blotting, polymerase chain reaction (PCR), and quantitative polymerase chain reaction (qPCR). Epithelial-mesenchymal transition (EMT) was induced in DF-HERSCs by treatment with transforming growth factor-β (TGF-β) or fetal bovine serum (FBS)-added medium. Characteristics of DF-HERSCs were compared with normal human oral keratinocytes (NHOKs) and normal human epidermal keratinocytes (NHEKs). Osteogenic differentiation and mineralization of DF-HERSCs were analyzed by alkaline phosphatase (ALP) and Alizarin red staining. All experiments were conducted in triplicate. RESULTS: Primary DF-HERSCs were isolated from DF. Epithelial phenotypes of DF-HERSCs were confirmed by morphological and Western blot analysis. PCR results demonstrated that the origin of DF-HERSCs was neither endothelial nor hematopoietic. Enamel matrix derivative (EMD)-associated genes were not expressed in DF-HERSCs. Treatment with TGF-β and FBS-added medium triggered the progression of EMT in DF-HERSCs. The acquired potency of differentiation and mineralization was shown in EMT-progressed DF-HERSCs. CONCLUSIONS: DF contains putative populations of HERSC, named DF-HERSC. DF-HERSCs shared common characteristics with NHOKs and NHEKs.
OBJECTIVES: The aim of this study was the isolation and characterization of dental follicle-derived Hertwig's epithelial root sheath cells (DF-HERSCs). MATERIALS AND METHODS: DF-HERSCs were isolated from dental follicle (DF)-derived single-cell suspensions. Their epithelial phenotypes were analyzed by Western blotting, polymerase chain reaction (PCR), and quantitative polymerase chain reaction (qPCR). Epithelial-mesenchymal transition (EMT) was induced in DF-HERSCs by treatment with transforming growth factor-β (TGF-β) or fetal bovine serum (FBS)-added medium. Characteristics of DF-HERSCs were compared with normal human oral keratinocytes (NHOKs) and normal human epidermal keratinocytes (NHEKs). Osteogenic differentiation and mineralization of DF-HERSCs were analyzed by alkaline phosphatase (ALP) and Alizarin red staining. All experiments were conducted in triplicate. RESULTS: Primary DF-HERSCs were isolated from DF. Epithelial phenotypes of DF-HERSCs were confirmed by morphological and Western blot analysis. PCR results demonstrated that the origin of DF-HERSCs was neither endothelial nor hematopoietic. Enamel matrix derivative (EMD)-associated genes were not expressed in DF-HERSCs. Treatment with TGF-β and FBS-added medium triggered the progression of EMT in DF-HERSCs. The acquired potency of differentiation and mineralization was shown in EMT-progressed DF-HERSCs. CONCLUSIONS: DF contains putative populations of HERSC, named DF-HERSC. DF-HERSCs shared common characteristics with NHOKs and NHEKs.
Authors: Emi Shimizu; Domenico Ricucci; Jeffrey Albert; Adel S Alobaid; Jennifer L Gibbs; George T-J Huang; Louis M Lin Journal: J Endod Date: 2013-05-22 Impact factor: 4.171
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