Immortalized cementoblasts and periodontal ligament cells in culture

Cementum, a mineralized tissue lining the surface of the tooth root, is required for formation of a functional periodontal ligament attachment during development. Additionally, during regeneration of tissues after disease, cementum is thought to play a critical role in the reparative process. Resear...

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Veröffentlicht in:	Bone (New York, N.Y.) Jg. 25; H. 1; S. 39 - 47
Hauptverfasser:	D’Errico, J.A, Ouyang, H, Berry, J.E, MacNeil, R.L, Strayhorn, C, Imperiale, M.J, Harris, N.L, Goldberg, H, Somerman, M.J
Format:	Journal Article Tagungsbericht
Sprache:	Englisch
Veröffentlicht:	New York, NY Elsevier Inc 01.07.1999 Elsevier Science
Schlagworte:	Animals Biological and medical sciences Bone sialoprotein Cell Survival - physiology Cells, Cultured Cementoblast Cementum Dental Cementum - physiology Female Fundamental and applied biological sciences. Psychology Male Mice Mice, Transgenic Mouth. Exocrine and endocrine salivary glands. Teeth. Esophagus Osteoblast Osteocalcin Osteopontin Periodontal Ligament - cytology Phenotype Reverse Transcriptase Polymerase Chain Reaction Vertebrates: digestive system Osteopontin Osteoblast Bone sialoprotein Osteocalcin Cementoblast Cementum Rodentia Cell biology ENT Experimental study Regeneration Vertebrata Periodontal ligament Mammalia Sialoproteins Synthesis Mouse Animal Established cell line Tooth
ISSN:	8756-3282, 1873-2763
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Abstract	Cementum, a mineralized tissue lining the surface of the tooth root, is required for formation of a functional periodontal ligament attachment during development. Additionally, during regeneration of tissues after disease, cementum is thought to play a critical role in the reparative process. Research efforts aimed toward understanding mechanisms involved in periodontal development and regeneration, and in particular the formation of root cementum, have been hampered by an inability to isolate and culture cells involved in cementum production, i.e., cementoblasts. Using classical techniques for osteoblast isolation, immortalized, heterogeneous cementoblast/periodontal ligament cell (CM/PDL) populations were established from cells lining the tooth root surface of: 1) CD-1 mice, where cells were immortalized using SV40, or 2) H-2K btsA58 “immorto” mice, where cells containing an immortalizing transgene were removed and cultured. CM/PDL populations were derived from tissues adherent to developing tooth root surfaces, while tissues adherent to the surrounding alveolar bone were specifically excluded from the population. Immortalized CM/PDL cells were characterized to ensure their phenotype reflected that previously demonstrated in situ and in primary, nonimmortalized cultures. Proteins/mRNAs associated with bone/cementum and known to be expressed by root lining cementoblasts, but not by PDL cells, in situ, e.g., bone sialoprotein, osteopontin, and osteocalcin, were expressed by cells within the immortalized populations. Furthermore, CM/PDL cells, in vitro, attached to bone sialoprotein in an arginine-glycine-aspartic acid (RGD)-dependent manner, promoted mineral nodule formation and exhibited a PTH/PTHrP-mediated cAMP response. These immortalized heterogeneous populations, containing both CM and PDL cells, provide a unique opportunity to study cells involved in cementogenesis and to enhance our knowledge of the mechanisms controlling development, maintenance, and regeneration of periodontal tissues.
AbstractList	Cementum, a mineralized tissue lining the surface of the tooth root, is required for formation of a functional periodontal ligament attachment during development. Additionally, during regeneration of tissues after disease, cementum is thought to play a critical role in the reparative process. Research efforts aimed toward understanding mechanisms involved in periodontal development and regeneration, and in particular the formation of root cementum, have been hampered by an inability to isolate and culture cells involved in cementum production, i.e., cementoblasts. Using classical techniques for osteoblast isolation, immortalized, heterogeneous cementoblast/periodontal ligament cell (CM/PDL) populations were established from cells lining the tooth root surface of: 1) CD-1 mice, where cells were immortalized using SV40, or 2) H-2K btsA58 “immorto” mice, where cells containing an immortalizing transgene were removed and cultured. CM/PDL populations were derived from tissues adherent to developing tooth root surfaces, while tissues adherent to the surrounding alveolar bone were specifically excluded from the population. Immortalized CM/PDL cells were characterized to ensure their phenotype reflected that previously demonstrated in situ and in primary, nonimmortalized cultures. Proteins/mRNAs associated with bone/cementum and known to be expressed by root lining cementoblasts, but not by PDL cells, in situ, e.g., bone sialoprotein, osteopontin, and osteocalcin, were expressed by cells within the immortalized populations. Furthermore, CM/PDL cells, in vitro, attached to bone sialoprotein in an arginine-glycine-aspartic acid (RGD)-dependent manner, promoted mineral nodule formation and exhibited a PTH/PTHrP-mediated cAMP response. These immortalized heterogeneous populations, containing both CM and PDL cells, provide a unique opportunity to study cells involved in cementogenesis and to enhance our knowledge of the mechanisms controlling development, maintenance, and regeneration of periodontal tissues. Cementum, a mineralized tissue lining the surface of the tooth root, is required for formation of a functional periodontal ligament attachment during development. Additionally, during regeneration of tissues after disease, cementum is thought to play a critical role in the reparative process. Research efforts aimed toward understanding mechanisms involved in periodontal development and regeneration, and in particular the formation of root cementum, have been hampered by an inability to isolate and culture cells involved in cementum production, i.e., cementoblasts. Using classical techniques for osteoblast isolation, immortalized, heterogeneous cementoblast/periodontal ligament cell (CM/PDL) populations were established from cells lining the tooth root surface of: 1) CD-1 mice, where cells were immortalized using SV40, or 2) H-2KbtsA58 "immorto" mice, where cells containing an immortalizing transgene were removed and cultured. CM/PDL populations were derived from tissues adherent to developing tooth root surfaces, while tissues adherent to the surrounding alveolar bone were specifically excluded from the population. Immortalized CM/PDL cells were characterized to ensure their phenotype reflected that previously demonstrated in situ and in primary, nonimmortalized cultures. Proteins/mRNAs associated with bone/cementum and known to be expressed by root lining cementoblasts, but not by PDL cells, in situ, e.g., bone sialoprotein, osteopontin, and osteocalcin, were expressed by cells within the immortalized populations. Furthermore, CM/PDL cells, in vitro, attached to bone sialoprotein in an arginine-glycineaspartic acid (RGD)-dependent manner, promoted mineral nodule formation and exhibited a PTH/PTHrP-mediated cAMP response. These immortalized heterogeneous populations, containing both CM and PDL cells, provide a unique opportunity to study cells involved in cementogenesis and to enhance our knowledge of the mechanisms controlling development, maintenance, and regeneration of periodontal tissues.Cementum, a mineralized tissue lining the surface of the tooth root, is required for formation of a functional periodontal ligament attachment during development. Additionally, during regeneration of tissues after disease, cementum is thought to play a critical role in the reparative process. Research efforts aimed toward understanding mechanisms involved in periodontal development and regeneration, and in particular the formation of root cementum, have been hampered by an inability to isolate and culture cells involved in cementum production, i.e., cementoblasts. Using classical techniques for osteoblast isolation, immortalized, heterogeneous cementoblast/periodontal ligament cell (CM/PDL) populations were established from cells lining the tooth root surface of: 1) CD-1 mice, where cells were immortalized using SV40, or 2) H-2KbtsA58 "immorto" mice, where cells containing an immortalizing transgene were removed and cultured. CM/PDL populations were derived from tissues adherent to developing tooth root surfaces, while tissues adherent to the surrounding alveolar bone were specifically excluded from the population. Immortalized CM/PDL cells were characterized to ensure their phenotype reflected that previously demonstrated in situ and in primary, nonimmortalized cultures. Proteins/mRNAs associated with bone/cementum and known to be expressed by root lining cementoblasts, but not by PDL cells, in situ, e.g., bone sialoprotein, osteopontin, and osteocalcin, were expressed by cells within the immortalized populations. Furthermore, CM/PDL cells, in vitro, attached to bone sialoprotein in an arginine-glycineaspartic acid (RGD)-dependent manner, promoted mineral nodule formation and exhibited a PTH/PTHrP-mediated cAMP response. These immortalized heterogeneous populations, containing both CM and PDL cells, provide a unique opportunity to study cells involved in cementogenesis and to enhance our knowledge of the mechanisms controlling development, maintenance, and regeneration of periodontal tissues. Cementum, a mineralized tissue lining the surface of the tooth root, is required for formation of a functional periodontal ligament attachment during development. Additionally, during regeneration of tissues after disease, cementum is thought to play a critical role in the reparative process. Research efforts aimed toward understanding mechanisms involved in periodontal development and regeneration, and in particular the formation of root cementum, have been hampered by an inability to isolate and culture cells involved in cementum production, i.e., cementoblasts. Using classical techniques for osteoblast isolation, immortalized, heterogeneous cementoblast/periodontal ligament cell (CM/PDL) populations were established from cells lining the tooth root surface of: 1) CD-1 mice, where cells were immortalized using SV40, or 2) H-2KbtsA58 "immorto" mice, where cells containing an immortalizing transgene were removed and cultured. CM/PDL populations were derived from tissues adherent to developing tooth root surfaces, while tissues adherent to the surrounding alveolar bone were specifically excluded from the population. Immortalized CM/PDL cells were characterized to ensure their phenotype reflected that previously demonstrated in situ and in primary, nonimmortalized cultures. Proteins/mRNAs associated with bone/cementum and known to be expressed by root lining cementoblasts, but not by PDL cells, in situ, e.g., bone sialoprotein, osteopontin, and osteocalcin, were expressed by cells within the immortalized populations. Furthermore, CM/PDL cells, in vitro, attached to bone sialoprotein in an arginine-glycineaspartic acid (RGD)-dependent manner, promoted mineral nodule formation and exhibited a PTH/PTHrP-mediated cAMP response. These immortalized heterogeneous populations, containing both CM and PDL cells, provide a unique opportunity to study cells involved in cementogenesis and to enhance our knowledge of the mechanisms controlling development, maintenance, and regeneration of periodontal tissues.
Author	Ouyang, H MacNeil, R.L Goldberg, H Imperiale, M.J Berry, J.E Strayhorn, C Harris, N.L D’Errico, J.A Somerman, M.J
Author_xml	– sequence: 1 givenname: J.A surname: D’Errico fullname: D’Errico, J.A email: somerman@umich.edu organization: Department of Periodontics/Prevention/Geriatrics, University of Michigan, Ann Arbor, MI, USA – sequence: 2 givenname: H surname: Ouyang fullname: Ouyang, H organization: Department of Periodontics/Prevention/Geriatrics, University of Michigan, Ann Arbor, MI, USA – sequence: 3 givenname: J.E surname: Berry fullname: Berry, J.E organization: Department of Periodontics/Prevention/Geriatrics, University of Michigan, Ann Arbor, MI, USA – sequence: 4 givenname: R.L surname: MacNeil fullname: MacNeil, R.L organization: Academic Affairs, School of Dental Medicine, University of Connecticut Health Center, Farmington, CT, USA – sequence: 5 givenname: C surname: Strayhorn fullname: Strayhorn, C organization: Department of Periodontics/Prevention/Geriatrics, University of Michigan, Ann Arbor, MI, USA – sequence: 6 givenname: M.J surname: Imperiale fullname: Imperiale, M.J organization: Department of Microbiology and Immunology, University of Michigan, Ann Arbor, MI, USA – sequence: 7 givenname: N.L surname: Harris fullname: Harris, N.L organization: School of Dentistry, Department of Biochemistry, The University of Western Ontario, London, Ontario, Canada – sequence: 8 givenname: H surname: Goldberg fullname: Goldberg, H organization: School of Dentistry, Department of Biochemistry, The University of Western Ontario, London, Ontario, Canada – sequence: 9 givenname: M.J surname: Somerman fullname: Somerman, M.J organization: Department of Periodontics/Prevention/Geriatrics, University of Michigan, Ann Arbor, MI, USA
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Keywords	Osteopontin Osteoblast Bone sialoprotein Osteocalcin Cementoblast Cementum Rodentia Cell biology ENT Experimental study Regeneration Vertebrata Periodontal ligament Mammalia Sialoproteins Synthesis Mouse Animal Established cell line Tooth
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Snippet	Cementum, a mineralized tissue lining the surface of the tooth root, is required for formation of a functional periodontal ligament attachment during...
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SubjectTerms	Animals Biological and medical sciences Bone sialoprotein Cell Survival - physiology Cells, Cultured Cementoblast Cementum Dental Cementum - physiology Female Fundamental and applied biological sciences. Psychology Male Mice Mice, Transgenic Mouth. Exocrine and endocrine salivary glands. Teeth. Esophagus Osteoblast Osteocalcin Osteopontin Periodontal Ligament - cytology Phenotype Reverse Transcriptase Polymerase Chain Reaction Vertebrates: digestive system
Title	Immortalized cementoblasts and periodontal ligament cells in culture
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