Characterization and Genetic Profiling of the Primary Cells and Tissues from Mandible of Mouse Fetus and Neonate
2
조회
0
다운로드
0
인용
0
참고문헌
키워드
초록
The stem cell research is emerging as a cutting edge topic for a new treatment for many chronic diseases. Recently, dental stem cell would be possible for regeneration of tooth itself as well as periodontal tissue. However, the study of the cell characterization is scarce. Therefore, we performed the genetic profiling and the characterization of mouse fetus/ neonate derived dental tissue and cell to find the identification during dental development. We separated dental arch from mandibles of 14.5 d fetal mice and neonate 0 d under the stereoscope, and isolated dental cells primarily from the tissues. Then, we examined morphology and the gene expression profiles of the primary cells and dental tissues from fetus/neonate and adult with RT-PCR. Primary dental cells showed heterogeneous but the majority was shown as fibroblast-like morphology. The change of population doubling time levels (PDLs) showed that the primary dental cells have growth potential and could be expanded under our culture conditions without reduction of growth rate. Immunocytochemical and flow cytometric analyses were performed to characterize the primary dental cell populations from both of fetus (E14.5) and neonate. Alpha smooth muscle actin (α-SMA), vimentin, and von Willebrand factor showed strong expression, but desmin positive cells were not detected in the primary dental cells. Most of the markers were not uniformly expressed, but found in subsets of cells, indicating that the primary dental cell population is heterogeneous, and characteristics of the populations were changed during culture period. And mesenchymal stem cell markers were highly expressed. Gene expression profile showed Wnt family and its related signaling molecules, growth factors, transcription factors and tooth specific molecules were expressed both fetal and neonatal tissue. The tooth specific genes (enamelin, amelogenin, and DSPP) only expressed in neonate and adult stage. These expression patterns appeared same as primary fetal and neonatal cells. In this study we isolated primary cells from whole mandible of fetal and neonatal mice. And we investigated the characteristics of the primary cells and the profile of gene expressions, which are involved in epithelial–mesenchymal interactions during tooth development. Taken together, the primary dental cells in early passages or fetal and neonatal mandibles could be useful stem cell resources.
Abstract
The stem cell research is emerging as a cutting edge topic for a new treatment for many chronic diseases. Recently, dental stem cell would be possible for regeneration of tooth itself as well as periodontal tissue. However, the study of the cell characterization is scarce. Therefore, we performed the genetic profiling and the characterization of mouse fetus/ neonate derived dental tissue and cell to find the identification during dental development. We separated dental arch from mandibles of 14.5 d fetal mice and neonate 0 d under the stereoscope, and isolated dental cells primarily from the tissues. Then, we examined morphology and the gene expression profiles of the primary cells and dental tissues from fetus/neonate and adult with RT-PCR. Primary dental cells showed heterogeneous but the majority was shown as fibroblast-like morphology. The change of population doubling time levels (PDLs) showed that the primary dental cells have growth potential and could be expanded under our culture conditions without reduction of growth rate. Immunocytochemical and flow cytometric analyses were performed to characterize the primary dental cell populations from both of fetus (E14.5) and neonate. Alpha smooth muscle actin (α-SMA), vimentin, and von Willebrand factor showed strong expression, but desmin positive cells were not detected in the primary dental cells. Most of the markers were not uniformly expressed, but found in subsets of cells, indicating that the primary dental cell population is heterogeneous, and characteristics of the populations were changed during culture period. And mesenchymal stem cell markers were highly expressed. Gene expression profile showed Wnt family and its related signaling molecules, growth factors, transcription factors and tooth specific molecules were expressed both fetal and neonatal tissue. The tooth specific genes (enamelin, amelogenin, and DSPP) only expressed in neonate and adult stage. These expression patterns appeared same as primary fetal and neonatal cells. In this study we isolated primary cells from whole mandible of fetal and neonatal mice. And we investigated the characteristics of the primary cells and the profile of gene expressions, which are involved in epithelial–mesenchymal interactions during tooth development. Taken together, the primary dental cells in early passages or fetal and neonatal mandibles could be useful stem cell resources.
참고문헌 (41)
- Azizi, S.A, "Engraftment and migration of human bone marrow stromal cells implanted in the brains of albino rats--similarities to astrocyte grafts", 95, pp.3908~3913 (1998)
- Bianco, P, "nessuno e centomila: searching for the identity of mesodermal progenitors", 251, pp.257~263 (1999)
- Bianco, P, "Bone marrow stromal stem cels: nature, biology, and potential applications.", 19, pp.180~192 (2001)
- Blau, H. M, "The evolving concept of a stem cell : entity or function?", 105, pp.829~841 (2001)
- Cheng, "Differentiation of human bone marrow osteogenic stromal cels in vitro induction of the osteoblast phenotype by dexamethasone", endocri(logy 134), pp.277~86 (1994)
- Couble, M.L, "Odontoblast diferentiation of human dental pulp cels in explant cultures", 66, pp.129~138 (2000)
- Dassule, H.R, "Analysis of epithelial-mesenchymal interactions in the initial morphogenesis of the mamalian tooth", 202, pp.215~227 (1998)
- Doherty, M.J, "Vascular pericytes express osteogenic potential in vitro and in vivo", 13, pp.828~838 (1998)
- Fuchs, E, "Stem cells: a new lease on life", 100, pp.143~155 (2000)
- Gould, "Migration and division of progenitor cell populations in periodontal ligament after wounding" (1980)
- Gronthos, S, "Stem cell properties of human dental pulp stem cels", 81, pp.531~535 (2002)
- Graves, "The STRO-1+ fraction of adult human bone marrow contains the osteogenic precursors", pp.4164~73 (1994)
- Gronthos, S, "Postnatal human dental pulp stem cells (DPSCs) in vitro and in vivo", 97, pp.13625~13630 (2000)
- Hardy, "The secret life of the hair follicle", pp.55~61 (1992)
- Davidson, "Expresion of a human homeobox-containing gene is regulated by 1 2D3 in bone cells", pp.11~6 (1993)
- Isaka, J, "Participation of periodontal ligament cells with regeneration of alveolar bone", 72, pp.31423~ (2001)
- Jahoda, "Induction of follicle formation and hair growth by vibrisa dermal papilae implanted into rat ear wounds", pp.1103~115 9 (1992)
- Jernvall, J, "Reiterative signaling and patterning during mamalian tooth morphogenesis", 92, pp.19~29 (2000)
- Kettunen, P.Karavanova, I.Theslef, I, "Responsiveness of developing dental tisues to fibroblast growth factors expression of splicing alternatives of FGFR1, -2, -3, and of FGFR4 ; and stimulation of cell proliferation by FGF-2, -4, - 8, and -9", 22, pp.374~385 (1998)
- Kollar, "The induction of hair follicles by embryonic dermal papillae", pp.374~8 (1970)
- Kollar, "Tisue interactions in embryonic mouse toth germs Reorganization of the dental epithelium during tooth-germ reconstruction", pp.159~71 (1970)
- Kollar, "Tisue interactions in embryonic mouse tooth germs The inductive role of the dental papilla", pp.173~86 (1970)
- Lekic, P, "Phenotypic comparison of periodontal ligament cells in vivo and in vitro", 36, pp.71~79 (2001)
- Lumsden, "Spatial organization of the epithelium and the role of neural crest cels in the initiation of the mamalian tooth germ", pp.103 155~69 (1988)
- Maas, R, "The genetic control of early tooth development", 8, pp.4~39 (1997)
- Mitsiadis, T.A, "Mouse Serrate-1 (Jagged-1): expression in the developing tooth is regulated by epithelial-mesenchymal interactions and fibroblast growth factor-4", 124, pp.1473~1483 (1997)
- Mitsiadis, T.A, "Delta-notch signaling in odontogenesis : correlation with cytodiferentiation and evidence for feedback regulation", 204, pp.420~431 (1998)
- Mitsiadis, T.A, "Expresion of Notch 1, 2 and 3 is regulated by epithelial- mesenchymal interactions and retinoic acid in the developing mouse tooth and associated with determination of ameloblast cell fate", 130, pp.407~418 (1995)
- Miura, M, "SHED: stem cells from human exfoliated deciduous teth", 100, pp.5807~5812 (2003)
- Nakashima, M,, "Induction of reparative dentin formation by ultrasound-mediated gene delivery of growth", differentiation factor 11 Stimulation of proliferation and differentiation of dog dental pulp cells in serum-free culture medium by insulin-like growth factor 11, 2003, pp.14, 591~597
- Peters, H, "Teeth. Where and how to make them", 15, pp.59~65 (1999)
- Prockop, D.J, "Marow stromal cells as stem cells for nonhematopoietic tissues", 276, pp.71~74 (1997)
- Tissue Int, "Efects of parathyroid hormone on odontogenesis of the mouse embryonic molar tooth in vitro" (1987)
- Seo, B.M, "Investigation of multipotent postnatal stem cells from human periodontal ligament", 364, pp.149~155 (2004)
- Shiba, H, "Differential effects of various growth factors and cytokines on the syntheses of DNA , type I collagen, laminin, fibronectin, osteonectin", secreted protein, acidic and rich in cysteine (SPARC), and alkaline phosphatase by human pulp cells in culture, 1998, pp.174, 194~205
- Theslef, I.Sharpe, P, "Signalling networks regulating dental development.", 67, pp.111~123 (1997)
- Theslef, I, "Epithelial-mesenchymal signalling regulating tooth morphogenesis", 116, pp.1647~1648 (2003)
- Tucker, A.S, "Molecular genetics of tooth morphogenesis and patterning the right shape in the right place", 78, pp.826~834 (1999)
- Weiss, K, "Perspectives on genetic aspects of dental patterning", 106, pp.55~63 (1998)
- Weiss, K.M, "Dynamic interactions and the evolutionary genetics of dental patterning", 9, pp.369~398 (1998)
- Onishi, T, "Stimulation of proliferation and differentiation of dog dental pulp cells in serum-free culture medium by insulin-like growth factor", 44, pp.361~371 (1999)