Isolating mineralized bone and bone marrow mRNA from transiliac bone biopsies stored in a stabilizing solution: A comparative study

The molecular mechanisms underlying metabolic bone diseases, including renal osteodystrophy, are poorly understood. Transcriptomics are increasingly used to characterize biological molecular networks and prove promising in identifying therapeutic targets and biomarkers. A reliable method for obtaini...

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Bibliographic Details
Published in:Bone Reports 2022-12, Vol.17, p.101624-101624, Article 101624
Main Authors: de Loor, Henriette, Smout, Dieter, Jørgensen, Hanne, Meng, Catarina, Van Craenenbroeck, Amaryllis H., Evenepoel, Pieter
Format: Article
Language:English
Subjects:
Bone
Bone marrow
Full Length
Human
mRNA
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Summary:The molecular mechanisms underlying metabolic bone diseases, including renal osteodystrophy, are poorly understood. Transcriptomics are increasingly used to characterize biological molecular networks and prove promising in identifying therapeutic targets and biomarkers. A reliable method for obtaining sufficient amounts of high quality RNA from human bone biopsies is a prerequisite for the implementation of molecular diagnostics in clinical research and practice. The present study aimed to develop a simple and adequate method for isolating bone and bone marrow mRNA from transiliac bone biopsies. Several storage, separation, and extraction procedures were compared. The procedure was optimized in pig samples and subsequently validated in human samples. Appropriate amounts of mineralized bone and bone marrow mRNA of moderate to high quality were obtained from transiliac bone biopsies that were immersed in the stabilizing solution Allprotect Tissue Reagent at room temperature for up to 3 days prior to freezing. After thawing, bone marrow and mineralized bone were separated by a multistep centrifugation procedure and subsequently disrupted and homogenized by a bead crusher. Appropriate separation of mineralized bone and bone marrow was confirmed by discriminatory gene expression profiles. [Display omitted] •Molecular diagnostics increasingly gain interest in clinical practice.•A bone biopsy immersed in a stabilization reagent yields moderate mRNA quality.•Use of a stabilization reagent allows for easy separation of bone and bone marrow.
ISSN:2352-1872
2352-1872
DOI:10.1016/j.bonr.2022.101624