Validation of the Reference Genes for the Gene Expression Studies in Different Cell Lines of Pig

Biomed Res Int. 2021 Aug 18:2021:5364190. doi: 10.1155/2021/5364190. eCollection 2021.

Abstract

Reverse transcription quantitative real-time polymerase chain reaction is one of the important methods to investigate gene expression in cells and tissues. However, if the data cannot be normalized with appropriate reference genes, the results may be unreliable. In this study, we detected the expression of 15 reference genes in three pig cell lines. The results showed that SDHA and ALDOA were the most stable reference genes in 3D4/21 cells. TOP2B, TBP, and PPIA were the most stable reference genes in PK-15 cells. SDHA and ALDOA were the most stable reference genes in IPEC-J2 cells. In addition, each cell line only needs to use two reference genes to standardize the expression of target genes. Taken together, this study provides a reference for different pig cell lines to select reference genes and also provides a theoretical basis for the use of these cell lines in related functional researches.

Publication types

  • Validation Study

MeSH terms

  • Animals
  • Biomarkers / metabolism
  • Cell Line
  • Gene Expression Profiling / methods
  • Gene Expression Profiling / standards*
  • RNA, Messenger / genetics*
  • RNA, Messenger / metabolism
  • Real-Time Polymerase Chain Reaction / methods*
  • Reference Standards
  • Swine

Substances

  • Biomarkers
  • RNA, Messenger