An improved method to isolate primary human osteocytes from bone
-
Anne Bernhardt
,
Sophie Wolf
Abstract
Osteocytes are of high importance in bone metabolism as they orchestrate bone remodeling, react to mechanosensory stimuli and have endocrine functions. In vitro investigations with osteocytes are therefore of high relevance for biomaterial and drug testing. The application of primary human cells instead of rodent osteocyte cell lines like MLOY4 and IDG SW3 is desirable but provides the challenge of isolating these cells, which are deeply embedded into the mineralized bone matrix. The present study describes an improved protocol for the isolation of human primary osteocytes. In contrast to an already established protocol, resting steps between the demineralization /digestion steps of the bone particles considerably improved the yield of osteocytes. Real-time polymerase chain reaction (PCR) analysis revealed the expression of typical osteocyte markers like osteocalcin, E11/podoplanin and dentin matrix protein 1 (DMP-1).
Funding source: German Research Foundation (DFG)
Award Identifier / Grant number: BE 5139/3-1
Funding statement: This study was funded by the German Research Foundation (DFG), (funder Id: http://dx.doi.org/10.13039/501100001659, BE 5139/3-1).
Author Statement
Conflict of interest: Authors state no conflict of interest.
Informed consent: Informed consent is not applicable.
Ethical approval: The conducted research is not related to either human or animal use.
References
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Articles in the same Issue
- Frontmatter
- Review
- Radioguided surgery: physical principles and an update on technological developments
- Research articles
- EOG-based eye movement recognition using GWO-NN optimization
- Effective brain connectivity estimation between active brain regions in autism using the dual Kalman-based method
- Epileptic seizure detection on EEG signals using machine learning techniques and advanced preprocessing methods
- Pattern recognition of head movement based on mechanomyography and its application
- A novel approach to the diagnostic assessment of carpal tunnel syndrome based on the frequency domain of the compound muscle action potential
- Automatic hypernasality grade assessment in cleft palate speech based on the spectral envelope method
- Multimodal feature learning and fusion on B-mode ultrasonography and sonoelastography using point-wise gated deep networks for prostate cancer diagnosis
- Automatic analysis of image quality by ETR-1 for the On-Board Imager
- An improved method to isolate primary human osteocytes from bone
- Experimental investigation on the effect of drill quality on the performance of bone drilling
