Validation of reference genes for RT-PCR in osteoblastic cells derived from rat bone marrow

• Grant number: 15/05376-4
• Support Opportunities: Scholarships in Brazil - Scientific Initiation
• Start date: June 01, 2015
• End date: July 25, 2016
• Field of knowledge: Health Sciences - Dentistry - Oral and Maxillofacial Surgery
• Principal Investigator: Adalberto Luiz Rosa
• Grantee: Jaqueline Isadora Reis Ramos
• Host Institution: Faculdade de Odontologia de Ribeirão Preto (FORP). Universidade de São Paulo (USP). Ribeirão Preto , SP, Brazil

Abstract

The Real-Time PCR assay is the first choice to evaluate the expression of genes involved in osteoblastic differentiation, thanks to its reproducibility. It request the normalization of the evaluated genes in relation to reference genes in order to minimize technical errors. For this, the reference genes should display stable expression along the multiple differentiation process. In keeping with this, the aim of this project is to evaluate the expression of 32 reference genes along the osteoblastic differentiation of rat bone marrow cells. The data of cycle threshold of all genes will be evaluated using the Expression Suit software to get a score meaning the gene stability along the differentiation process. Cell cultures will be monitored through cell proliferation and osteoblastic differentiation will be evaluated by expression of bone marker genes Runx2, alkaline phosphatase (ALP) and osteocalcin together with ALP activity and matrix mineralization. Data will be analyzed by ANOVA or Kruskall-Wallis when appropriated.

Scientific publications

(References retrieved automatically from Web of Science and SciELO through information on FAPESP grants and their corresponding numbers as mentioned in the publications by the authors)

ABUNA, RODRIGO P. F.; OLIVEIRA, FABIOLA S.; RAMOS, JAQUELINE I. R.; LOPES, HELENA B.; FREITAS, GILEADE P.; SOUZA, ALANN T. P.; BELOTI, MARCIO M.; ROSA, ADALBERTO L.. Selection of reference genes for quantitative real-time polymerase chain reaction studies in rat osteoblasts. Journal of Cellular Physiology, v. 234, n. 1, p. 749-756, JAN 2019. (15/05376-4)