RRC ID 45472
著者 Hashimoto A, Yamaguchi Y, Chiu LD, Morimoto C, Fujita K, Takedachi M, Kawata S, Murakami S, Tamiya E.
タイトル Time-lapse Raman imaging of osteoblast differentiation.
ジャーナル Sci Rep
Abstract Osteoblastic mineralization occurs during the early stages of bone formation. During this mineralization, hydroxyapatite (HA), a major component of bone, is synthesized, generating hard tissue. Many of the mechanisms driving biomineralization remain unclear because the traditional biochemical assays used to investigate them are destructive techniques incompatible with viable cells. To determine the temporal changes in mineralization-related biomolecules at mineralization spots, we performed time-lapse Raman imaging of mouse osteoblasts at a subcellular resolution throughout the mineralization process. Raman imaging enabled us to analyze the dynamics of the related biomolecules at mineralization spots throughout the entire process of mineralization. Here, we stimulated KUSA-A1 cells to differentiate into osteoblasts and conducted time-lapse Raman imaging on them every 4 hours for 24 hours, beginning 5 days after the stimulation. The HA and cytochrome c Raman bands were used as markers for osteoblastic mineralization and apoptosis. From the Raman images successfully acquired throughout the mineralization process, we found that β-carotene acts as a biomarker that indicates the initiation of osteoblastic mineralization. A fluctuation of cytochrome c concentration, which indicates cell apoptosis, was also observed during mineralization. We expect time-lapse Raman imaging to help us to further elucidate osteoblastic mineralization mechanisms that have previously been unobservable.
巻・号 5
ページ 12529
公開日 2015-7-27
DOI 10.1038/srep12529
PII srep12529
PMID 26211729
PMC PMC4515588
MeSH Animals Calcification, Physiologic / physiology* Cell Differentiation / physiology Cell Line Cytochromes c / metabolism Mice Microscopy / methods* Osteoblasts / cytology* Osteoblasts / physiology* Spectrum Analysis, Raman / methods* Time-Lapse Imaging / methods* beta Carotene / metabolism
IF 3.998
引用数 20
WOS 分野 MULTIDISCIPLINARY SCIENCES
リソース情報
ヒト・動物細胞 KUSA-A1(RCB2081)