Establishment and characterization of a spontaneously immortalized mouse ameloblast-lineage cell line

Nakata, Akira; Kameda, Takashi; Nagai, Hirokazu; Ikegami, Kenji; Duan, Yuqing; Terada, Kunihiko; Sugiyama, Toshihiro

doi:10.1016/s0006-291x(03)01467-0

Cited by 87 publications

(88 citation statements)

References 16 publications

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“…170-6527; Biorad, Mississauga, Ont., Canada), according to the manufacturer's instructions. Three different cell types, primary human gingival fibroblasts [Pender and McCulloch, 1991], mouse ameloblast-lineage cells [Nakata et al, 2003] and mouse MC3T3-E1 osteoblasts [Sudo et al, 1983] were grown under the described culture conditions to approximately 80% confluence in 100-mm tissue culture dishes, washed with PBS, harvested and dispersed into a single-cell suspension by treatment with 0.05% Trypsin/0.53 m M EDTA (Cat. No.…”

Section: Cell Attachment Assaysmentioning

confidence: 99%

Comparative Temporospatial Expression Profiling of Murine Amelotin Protein during Amelogenesis

Somogyi-Ganss

Nakayama

Iwasaki

et al. 2011

Cells Tissues Organs

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Tooth enamel is formed in a typical biomineralization process under the guidance of specific organic components. Amelotin (AMTN) is a recently identified, secreted protein that is transcribed predominantly during the maturation stage of enamel formation, but its protein expression profile throughout amelogenesis has not been described in detail. The main objective of this study was to define the spatiotemporal expression profile of AMTN during tooth development in comparison with other known enamel proteins. A peptide antibody against AMTN was raised in rabbits, affinity purified and used for immunohistochemical analyses on sagittal and transverse paraffin sections of decalcified mouse hemimandibles. The localization of AMTN was compared to that of known enamel proteins amelogenin, ameloblastin, enamelin, odontogenic ameloblast-associated/amyloid in Pindborg tumors and kallikrein 4. Three-dimensional images of AMTN localization in molars at selected ages were reconstructed from serial stained sections, and transmission electron microscopy was used for ultrastructural localization of AMTN. AMTN was detected in ameloblasts of molars in a transient fashion, declining at the time of tooth eruption. Prominent expression in maturation stage ameloblasts of the continuously erupting incisor persisted into adulthood. In contrast, amelogenin, ameloblastin and enamelin were predominantly found during the early secretory stage, while odontogenic ameloblast-associated/amyloid in Pindborg tumors and kallikrein 4 expression in maturation stage ameloblasts paralleled that of AMTN. Secreted AMTN was detected at the interface between ameloblasts and the mineralized enamel. Recombinant AMTN protein did not mediate cell attachment in vitro. These results suggest a primary role for AMTN in the late stages of enamel mineralization.

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Section: Cell Attachment Assaysmentioning

confidence: 99%

Comparative Temporospatial Expression Profiling of Murine Amelotin Protein during Amelogenesis

Somogyi-Ganss

Nakayama

Iwasaki

et al. 2011

Cells Tissues Organs

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“…Various cell lines have been established with SV40 transfection for investigating dental morphogenesis, such as cell lines from dental epithelium [29,30] and dental mesenchyme [31,32]. Dental papilla cells have the tendency to differentiate into odontoblasts.…”

Section: Discussionmentioning

confidence: 99%

Application of immortalized mouse dental papilla cells for tooth bioengineering

Epasinghe¹,

Kim²,

Wu³

et al. 2018

Dent Oral Craniofac Res

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Mouse dental papilla cells (MDPCs) can be differentiated into component cell types that have the ability to regenerate the pulpo-dentinal complex. At the cap stage of tooth development, MDPC component cells are programmed for tooth generation. Therefore, isolated and cultivated component cells can be useful in evaluating cellular, molecular, and environmental scenarios in tooth germ formation and tooth morphogenesis. Increasing the proliferation capacity of MDPCs and preserving their original phenotypic and genotypic characteristics for tooth organ regeneration are the main focus of this study. An immortalized mouse dental papilla cell line was created via the intracellular insertion of SV40 T antigens into the nucleus by lentivirus particles. The generated clonally isolated SV40 T immortalized MDPC line was then characterized and validated for transfection success and efficiency. These cells displayed a higher proliferation rate, and both genotype and phenotype characteristics were similar to those of the original primary cell line. These results were verified via the expression of a broad array of tooth-specific markers. Furthermore, the test results to show that transformed cells had preserved multi-potency were also positive. Thus, the stable immortalized MDPCs may be used to determine the mechanisms of an array of developmental phenomena, such as early dental cell proliferation, reconstitution of tooth germ, dentine mineralization and other significant growth factor signaling pathways influencing tooth morphogenesis.

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“…Epithelial Rests of Malassez (REM), present in mature human periodontal tissues, may be a suitable source for human dental epithelium [130]. In addition, current attempts to reprogram cell sources include the development of spontaneously derived immortalized murine dental epithelial cell lines, which when grown in low calcium-supplemented media, can express enamel markers such as amelogenin [131,132], and the use of single-cell suspensions generated from tooth germ tissue [133]. Lessons from these studies should assist efforts to generate suitable human dental epithelial cell lines from available tissues such as Hertwig's epithelial root sheath or epithelial cell REM.…”

Section: Unmet Needsmentioning

confidence: 99%