Generation and characterization of two immortalized human osteoblastic cell lines useful for epigenetic studies

Flor M. Pérez-Campo, Tobias May, Jeannette Zauers, Carolina Sañudo, Jesus Delgado-Calle, Jana Arozamena, María T. Berciano, Miguel Lafarga, José A. Riancho

Research output: Contribution to journalArticle

4 Citations (Scopus)

Abstract

Abstract: Different model systems using osteoblastic cell lines have been developed to help understand the process of bone formation. Here, we report the establishment of two human osteoblastic cell lines obtained from primary cultures upon transduction of immortalizing genes. The resulting cell lines had no major differences to their parental lines in their gene expression profiles. Similar to primary osteoblastic cells, osteocalcin transcription increased following 1,25-dihydroxyvitamin D3 treatment and the immortalized cells formed a mineralized matrix, as detected by Alizarin Red staining. Moreover, these human cell lines responded by upregulating ALPL gene expression after treatment with the demethylating agent 5-aza-2′-deoxycytidine (AzadC), as shown before for primary osteoblasts. We further demonstrate that these cell lines can differentiate in vivo, using a hydroxyapatite/tricalcium phosphate composite as a scaffold, to produce bone matrix. More importantly, we show that these cells respond to demethylating treatment, as shown by the increase in SOST mRNA levels, the gene encoding sclerostin, upon treatment of the recipient mice with AzadC. This also confirms, in vivo, the role of DNA methylation in the regulation of SOST expression previously shown in vitro. Altogether our results show that these immortalized cell lines constitute a particularly useful model system to obtain further insight into bone homeostasis, and particularly into the epigenetic mechanisms regulating sclerostin production.

Original languageEnglish (US)
Pages (from-to)1-11
Number of pages11
JournalJournal of Bone and Mineral Metabolism
DOIs
StateAccepted/In press - Apr 2 2016

Fingerprint

Epigenomics
Cell Line
decitabine
Deoxycytidine
Bone Matrix
Calcitriol
Osteocalcin
DNA Methylation
Durapatite
Osteoblasts
Transcriptome
Osteogenesis
Genes
Homeostasis
Staining and Labeling
Gene Expression
Bone and Bones
Messenger RNA

Keywords

  • DNA methylation
  • Epigenetics
  • Immortalized osteoblastic cell lines
  • Sclerostin
  • SOST

ASJC Scopus subject areas

  • Endocrinology
  • Endocrinology, Diabetes and Metabolism
  • Orthopedics and Sports Medicine

Cite this

Generation and characterization of two immortalized human osteoblastic cell lines useful for epigenetic studies. / Pérez-Campo, Flor M.; May, Tobias; Zauers, Jeannette; Sañudo, Carolina; Delgado-Calle, Jesus; Arozamena, Jana; Berciano, María T.; Lafarga, Miguel; Riancho, José A.

In: Journal of Bone and Mineral Metabolism, 02.04.2016, p. 1-11.

Research output: Contribution to journalArticle

Pérez-Campo, Flor M. ; May, Tobias ; Zauers, Jeannette ; Sañudo, Carolina ; Delgado-Calle, Jesus ; Arozamena, Jana ; Berciano, María T. ; Lafarga, Miguel ; Riancho, José A. / Generation and characterization of two immortalized human osteoblastic cell lines useful for epigenetic studies. In: Journal of Bone and Mineral Metabolism. 2016 ; pp. 1-11.
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