ArticleBMC biotechnology2021
Optimizations for identifying reference genes in bone and cartilage bioengineering.
Article in BMC biotechnology, 2021. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 7 papers.
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Who cites it
7 citing papers in PubMed.
- Recapitulation ofJournal of orthopaedic translation · 2025Article
- Reference Genes for Expression Analyses by qRT-PCR inMicroorganisms · 2024Article
- Selection of reference genes in liproxstatin-1-treated K562 Leukemia cells via RT-qPCR and RNA sequencing.Molecular biology reports · 2024Article
- Osteochondrogenesis by TGF-β3, BMP-2 and noggin growth factor combinations in anFrontiers in bioengineering and biotechnology · 2023Article
- Evaluation of candidate reference genes for quantitative real-time PCR normalization in blood from red deer developing antlers.Scientific reports · 2022Article
- Article
- Circulating MicroRNAs as Non-invasive Biomarkers for Canine Cushing's Syndrome.Frontiers in veterinary science · 2021Article
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Authors and funding
5 authors.
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Abstract
backgroundReverse transcription quantitative real-time polymerase chain reaction (RT-qPCR) remains one of the best-established techniques to assess gene expression patterns. However, appropriate reference gene(s) selection remains a critical and challenging subject in which inappropriate reference gene selction can distort results leading to false interpretations. To date, mixed opinions still exist in how to choose the most optimal reference gene sets in accodrance to the Minimum Information for Publication of Quantitative Real-Time PCR Experiments (MIQE) guideline. Therefore, the purpose of this study was to investigate which schemes were the most feasible for the identification of reference genes in a bone and cartilage bioengineering experimental setting. In this study, rat bone mesenchymal stem cells (rBMSCs), skeletal muscle tissue and adipose tissue were utilized, undergoing either chondrogenic or osteogenic induction, to investigate the optimal reference gene set identification scheme that would subsequently ensure stable and accurate interpretation of gene expression in bone and cartilage bioengineering.
resultsThe stability and pairwise variance of eight candidate reference genes were analyzed using geNorm. The V
conclusionsBased on these results, the proposed cut-off value of Vn/n + 1 under 0.15, according to the geNorm algorithm, should be considered with caution. For cell only experiments, at least rBMSCs, a Vn/n + 1 under 0.15 is sufficient in RT-qPCR studies. However, when using certain tissue types such as skeletal muscle and adipose tissue the minimum Vn/n + 1 should be used instead as this provides a far superior mode of generating accurate gene expression results. We thus recommended that when the stability and variation of a candidate reference genes in a specific study is unclear the minimum Vn/n + 1 should always be used as this ensures the best and most accurate gene expression value is achieved during RT-qPCR assays.
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