ArticleJOR spine2018
Isolation of high-quality RNA from intervertebral disc tissue via pronase predigestion and tissue pulverization.
Article in JOR spine, 2018. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 28 papers.
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Who cites it
28 citing papers in PubMed, 32 citations in OpenAlex.
- Organ-on-a-Chip Technology: Advances in Modeling Inflammation, Neurovascularization and Nociception in Human Intervertebral Disc Degeneration.Smart medicine · 2026Review
- ISSLS Prize in Basic Science 2026: Early markers of mechanical modulation in whole bovine intervertebral discs loaded in a multiaxial bioreactor.European spine journal : official publication of the European Spine Society, the European Spinal Deformity Society, and the European Section of the Cervical Spine Research Society · 2026Article
- TIE2-positive cells in the nucleus pulposus with a purpose: the who, what and why.Journal of biomedical science · 2026Review
- Combined Flexion, Torsion and Compression Drive Distinct Intervertebral Disc Failure Mechanisms Under Asymmetric, High-Cycle Loading.JOR spine · 2026Article
- Advancement in the Methods for Isolating High Quality RNA From Mouse and Rat Intervertebral Disc.JOR spine · 2026Article
- Glucose concentration modulates bovine intervertebral disc response under physiological loading.Frontiers in bioengineering and biotechnology · 2026Article
- A proteomics investigation of primary human articular chondrocyte isolation.Osteoarthritis and cartilage open · 2025Article
- Short-Term Dynamic Unloading of Bovine Tail Discs in Culture Partially Mitigates Induced Degeneration After One-Strike Trigger.JOR spine · 2025Article
- Multiaxial rotational loading compromises the transition zone of the intervertebral disc: Ex vivo study using next-generation bioreactors.Bioengineering & translational medicine · 2025Article
- Article
- Repetitive strikes loading organ culture model to investigate the biological and biomechanical responses of the intervertebral disc.JOR spine · 2024Article
- Dynamics of N6-methyladenosine modification during aging and their potential roles in the degeneration of intervertebral disc.JOR spine · 2024Article
- Silicone implant surface microtopography modulates inflammation and tissue repair in capsular fibrosis.Frontiers in immunology · 2024Article
- Harmonization and standardization of nucleus pulposus cell extraction and culture methods.JOR spine · 2023Article
- Identification of novel gene signatures and immune cell infiltration in intervertebral disc degeneration using bioinformatics analysis.Frontiers in molecular biosciences · 2023Article
- Chemonucleolysis combined with dynamic loading for inducing degeneration in bovine caudal intervertebral discs.Frontiers in bioengineering and biotechnology · 2023Article
- Raloxifene Stimulates Estrogen Signaling to Protect Against Age- and Sex-Related Intervertebral Disc Degeneration in Mice.Frontiers in bioengineering and biotechnology · 2022Article
- Integrative Bioinformatics Analysis Revealed Mitochondrial Dysfunction-Related Genes Underlying Intervertebral Disc Degeneration.Oxidative medicine and cellular longevity · 2022Article
- Transcriptional profiling of intervertebral disc in a post-traumatic early degeneration organ culture model.JOR spine · 2021Article
- A comprehensive tool box for large animal studies of intervertebral disc degeneration.JOR spine · 2021Article
Corrections and comments
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Authors and funding
6 authors at 1 institution in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
The isolation of high-quality RNA from the intervertebral disc and especially from the nucleus pulposus is challenging due to the low cellularity and high proteoglycan content of this tissue. In this study, we report a simple modification of the standard guanidinium thiocyanate-phenol-chloroform extraction method, which involves enzymatic predigestion of the tissue prior to standard RNA isolation. Yield, purity and integrity of RNA isolated from bovine nucleus pulposus, inner annulus fibrosus and outer annulus fibrosus were compared among complete matrix digestion, predigestion and pulverization, pulverization alone, and pulverization followed by on-column purification. With predigestion, the average yield of RNA obtained from bovine nucleus pulposus was 8.82 ± 2.05 ng/mg of wet tissue with 260/280 and 260/230 optical density ratios of 1.91 ± 0.15 and 1.84 ± 0.30, respectively. RIN analysis indicated that RNA quality was best preserved with the predigestion method (RNA integrity number > 7), and the extracted RNA was suitable for real-time polymerase chain reaction. This method is of importance for gene expression studies on intervertebral disc development, degeneration and repair, and we anticipate that it may be further applied to other tissues rich in proteoglycans.
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Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.