Identification of suitable reference genes for mesenchymal stem cells from menstrual blood of women with endometriosis

Autor(es)

Zucherato, Victoria S. ^[1] ; Penariol, Leticia B. C. ^[1] ; Silva, Lilian E. C. M. ^[2] ; Padovan, Cristiana C. ^[1] ; Poli-Neto, Omero B. ^[1] ; Rosa-e-Silva, Julio C. ^[1] ; Ferriani, Rui A. ^{[3, 1]} ; Meola, Juliana ^{[3, 1]}

Afiliação do(s) autor(es)

^[1] Univ Sao Paulo, Ribeirao Preto Med Sch, Dept Gynecol & Obstet, Clin Hosp, Bandeirantes Ave 3900, 8th Floor, Univ Campus, BR-14049900 Ribeirao Preto, SP - Brazil

^[2] Univ Sao Paulo, Ribeirao Preto Med Sch, Multiuser Lab, Ribeirao Preto, SP - Brazil

^[3] CNPq, Natl Inst Hormones & Womans Hlth, Porto Alegre, RS - Brazil

• Tipo de documento: Artigo Científico
• Fonte: SCIENTIFIC REPORTS; v. 11, n. 1 MAR 8 2021.
• Citações Web of Science: 0

Resumo

It has been suggested that menstrual blood-derived mesenchymal stem/stromal cells (MenMSCs) are associated with the etiopathogenesis of endometriosis and considerable effort has been invested in searching for target genes and deciphering associated molecular pathways. However, reference gene stability for proper reproducible normalization in the analyses of the expression data validation is still unexplored in this experimental context. Therefore, in this exploratory study, we used stringent case and control selection criteria and collected menstrual blood from women with a laparoscopic diagnosis of advanced endometriosis and from fertile women without endometriosis. We tested for the first time the stability of 32 candidate reference genes to achieve increased accuracy and reliable results in the quantification of gene expression and direct future experiments using reverse transcription-quantitative PCR (RT-qPCR) in MenMSCs for endometriosis studies. Using the RefFinder web tool, we recommend the EIF2B1 and POP4 reference genes for the normalization of RT-qPCR data in study designs similar to ours. Furthermore, we suggest avoiding the commonly used GAPDH and ACTB reference genes as they are unstable. This high-visibility study is capable of directing different experimental designs as MenMSCs are derived from a minimally invasive tissue source with multifunctional roles in regenerative medicine. (AU)

• Processo FAPESP: 13/22431-3 - Perfil diferencial de transcritos em células tronco mesenquimais obtidas do fluxo menstrual (MenMSCs) de mulheres com endometriose
• Beneficiário: Juliana Meola Lovato
• Modalidade de apoio: Auxílio à Pesquisa - Jovens Pesquisadores