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Selection and validation of reference genes by RT-qPCR for murine cementoblasts in mechanical loading experiments simulating orthodontic forces in vitro
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Selection and validation of reference genes by RT-qPCR for murine cementoblasts in mechanical loading experiments simulating orthodontic forces in vitro
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Selection and validation of reference genes by RT-qPCR for murine cementoblasts in mechanical loading experiments simulating orthodontic forces in vitro
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Journal Article
Selection and validation of reference genes by RT-qPCR for murine cementoblasts in mechanical loading experiments simulating orthodontic forces in vitro
2020
Overview
Different structures and cell types of the periodontium respond to orthodontic tooth movement (OTM) individually. Cementoblasts (OC/CM) located in the immediate vicinity of the fibroblasts on the cement have found way to the centre of actual research. Here, we identify and validate possible reference genes for OC/CM cells by RT-qPCR with and without static compressive loading. We investigated the suitability of 3 reference genes in an in vitro model of cementoblast cells using four different algorithms (Normfinder, geNorm, comparative delta-C
t
method and BestKeeper) under different confluences and time. Comparable to our previous publications about reference genes in OTM in rats and human periodontal ligament fibroblasts (hPDLF),
Rpl22
in murine OC/CM cells appears as the least regulated gene so that it represents the most appropriate reference gene. Furthermore, unlike to the expression of our recommended reference genes, the expression of additionally investigated target genes changes with confluence and under loading compression. Based on our findings for future RT-qPCR analyses in OC/CM cells,
Rpl22
or the combination
Rpl22/Tbp
should be favored as reference gene. According to our results, although many publications propose the use of
Gapdh
, it does not seem to be the most suitable approach.
Publisher
Nature Publishing Group UK,Nature Publishing Group,Nature Portfolio
Subject
/ 631/80
/ 692/4023
/ Animals
/ Dental Cementum - metabolism
/ Genes
/ Glyceraldehyde-3-phosphate dehydrogenase
/ Glyceraldehyde-3-Phosphate Dehydrogenase (Phosphorylating) - biosynthesis
/ Glyceraldehyde-3-Phosphate Dehydrogenase (Phosphorylating) - genetics
/ Humanities and Social Sciences
/ Mice
/ Periodontal Ligament - cytology
/ Real-Time Polymerase Chain Reaction
/ Ribosomal Proteins - biosynthesis
/ Ribosomal Proteins - genetics
/ RNA-Binding Proteins - biosynthesis
/ RNA-Binding Proteins - genetics
/ Science
/ TATA-Box Binding Protein - biosynthesis
/ TATA-Box Binding Protein - genetics
/ Teeth
