Identification of reference genes for quantitative PCR during C3H10T1/2 chondrogenic differentiation

• DOI: 10.1007/s11033-019-04713-x
• Publication Date: 2019-03-07T06:12:56Z
• Authors: Serena Cappato; Francesca Giacopelli; Laura Tonachini; Roberto Ravazzolo; Renata Bocciardi
• Keywords: Chondrogenesis; Reference Genes

C3H10T1/2, a mouse mesenchymal stem cell line, is well-known in vitro model of chondrogenesis that can be easily employed to recapitulate some the mechanisms intervening this process. Moreover, these cells used validate effect candidate molecules identified by high throughput screening approaches applied development targeted therapy for human disorders which chondrogenic differentiation may involved, as conditions characterized heterotopic endochondral bone formation. Chondrogenic C3H10T1/2 monitored applying quantitative polymerase chain reaction (qPCR), one most sensitive methods allows detection small dynamic changes gene expression between samples obtained under different experimental conditions. In work, we have qPCR monitor specific markers during micromass cultures. Then geNorm approach identify stable reference genes suitable get robust normalization data. Among 12 (Ap3d1, Csnk2a2, Cdc40, Fbxw2, Fbxo38, Htatsf1, Mon2, Pak1ip1, Zfp91, 18S, ActB, GAPDH) Mon2 and Ap3d1 ones chondrogenesis. GAPDH commonly literature, resulted an level too compared (Sox9, Collagen type 2a1, 10a1 1a1), therefore are actually less recommended conclusion, nine equally obtain variation differentiation.