Characterization of established cementoblast-like cell lines from human cementum-lining cells in vitro and in vivo.
To study cellular characteristics of human cementoblasts using a cellular model is important for understanding the mechanisms of homeostasis and regeneration of periodontal tissues. However, at present no immortalized human cementoblast cell line has been established due to limitation of the life span. In the present study, therefore, we attempted to establish human cementoblast-like cell lines by transfection with telomerase catalytic subunit hTERT gene. Two stable clones (HCEM-1 and -2) with high telomerase activity were obtained and they grew over 200 population doublings without significant growth retardation. The expression of mRNA for differentiation markers, type I collagen, alkaline phosphatase (ALP), runt-related transcription factor 2, osteocalcin, bone sialoprotein and cementum-derived protein was revealed in these clones by RT-PCR. Moreover, these cells showed high ALP activity and calcified nodule formation in vitro. Interestingly, HCEM-2 showed cementum like formation on the surface of hydroxyapatites granules by subcutaneous transplantation into immunodeficient mice with hydroxyapatite granules. Thus, we established human cementoblast-like cell lines. We suggest that HCEM cell lines can be useful cell models for investigating the characteristics of human cementoblasts.
Kitagawa
Masae
M
Department of Oral Maxillofacial Pathobiology, Hiroshima, 734-8553, Japan.
Tahara
Hidetoshi
H
Department of Cell and Molecular Biology, Graduate School of Biomedical Sciences, Hiroshima University, Hiroshima, 734-8553, Japan.
Kitagawa
Shoji
S
Department of Oral Maxillofacial Pathobiology, Hiroshima, 734-8553, Japan.
Oka
Hiroko
H
Department of Oral Maxillofacial Pathobiology, Hiroshima, 734-8553, Japan.
Kudo
Yasusei
Y
Department of Oral Maxillofacial Pathobiology, Hiroshima, 734-8553, Japan.
Sato
Sunao
S
Department of Oral Maxillofacial Pathobiology, Hiroshima, 734-8553, Japan.
Ogawa
Ikuko
I
Center of Oral Clinical Examination, Hiroshima University Hospital, Hiroshima, 734-8553, Japan.
Miyaichi
Mutsumi
M
Department of Oral Maxillofacial Pathobiology, Hiroshima, 734-8553, Japan.
Takata
Takashi
T
Department of Oral Maxillofacial Pathobiology, Hiroshima, 734-8553, Japan; Center of Oral Clinical Examination, Hiroshima University Hospital, Hiroshima, 734-8553, Japan. Electronic address: ttakata@hiroshima-u.ac.jp.
D000469:Alkaline Phosphatase / Q000235:genetics; D000818:Animals; D002460:Cell Line; D017690:Cell Transplantation / Q000379:methods; D024042:Collagen Type I / Q000235:genetics; D050659:Core Binding Factor Alpha 1 Subunit / Q000235:genetics; D003739:Dental Cementum / Q000166:cytology* / Q000378:metabolism / Q000637:transplantation; D015870:Gene Expression / Q000235:genetics; D006801:Humans; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D008819:Mice, Nude; D015675:Osteocalcin / Q000235:genetics; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014183:Transplantation, Heterologous
D024042:Collagen Type I; D050659:Core Binding Factor Alpha 1 Subunit; D012333:RNA, Messenger; C493709:RUNX2 protein, human; D015675:Osteocalcin; D000469:Alkaline Phosphatase