1. Odontoblast RNA stability in different temperature-based protocols for tooth storage.
- Author
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Conde MC, Nedel F, Campos VF, Smith AJ, Nör JE, Demarco FF, and Tarquinio SB
- Subjects
- Adolescent, Adult, Dental Pulp cytology, Dentin cytology, Electrophoresis, Agar Gel, Extracellular Matrix Proteins analysis, Glycoproteins analysis, Humans, Phosphoproteins analysis, RNA, Ribosomal, 18S analysis, RNA, Ribosomal, 28S analysis, Reverse Transcriptase Polymerase Chain Reaction, Sialoglycoproteins analysis, Young Adult, Cryopreservation methods, Odontoblasts cytology, RNA analysis, Tissue Preservation methods
- Abstract
Aim: To evaluate the effect of four tooth storage temperature-based methods on quality of RNA obtained from cells retrieved from human dental pulps and human pre-dentine., Methodology: RNA was isolated from dental pulp tissue and from cells retrieved by scraping the pre-dentine of freshly extracted human third molars (n = 15) using TRIzol(®) reagent. Teeth were randomly assigned to the following temperature conditions: immediate RNA isolation after tooth extraction, liquid nitrogen (24 h), -80 °C (24 h), 20 °C (24 h) and 4 °C (6 h). RNA integrity was checked by the density of 28S and 18S ribosomal RNA. RT-PCR was used to analyse the expression of odontoblast makers (DSPP, DMP1 and MEPE) and the housekeeping gene GAPDH., Results: All experimental conditions evaluated preserved RNA integrity. The three odontoblastic markers were amplified from the pulp tissue and from the cells associated with pre-dentine., Conclusion: The four storage options allowed RNA isolation for RT-PCR analysis. These findings may facilitate the use of clinically derived human dental pulp and odontoblasts for endodontic research., (© 2011 International Endodontic Journal.)
- Published
- 2012
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