Cell detachment rapidly induces changes in noncoding RNA expression in human mesenchymal stromal cells

BioTechniques

Elena Della Bella¹  Martin J Stoddart¹ 

[1] 1AO Research Institute, AO Foundation, Davos, Switzerland;

关键词: cell detachment;    circular RNA;    human mesenchymal stem cells;    loss of anchorage;    microRNA;    noncoding RNA;   

DOI  :  10.2144/btn-2019-0038

来源: DOAJ

【 摘 要 】

Aims: To identify differential expression of noncoding RNAs after trypsinization in human mesenchymal stromal cells (hMSCs), focusing on miRNAs, piRNAs and circRNAs. Methods: hMSCs from the bone marrow of three donors were collected for RNA extraction, either lysed directly in monolayer or trypsinized and lysed within 30 min. Total RNA was isolated and sequenced for the evaluation of miRNA and piRNA expression or RNaseR treated and labeled for circRNA array hybridization. RT-qPCR was performed to evaluate the stability of candidate reference genes. Results & conclusions: Alterations in levels of several noncoding RNAs are rapidly induced after trypsinization of hMSCs, affecting critical pathways. This should be carefully considered for a proper experimental design.

【 授权许可】

Unknown