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

Elena Della Bella | Zendy; Martin J. Stoddart | Zendy

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

Author(s) -

Elena Della Bella,

Martin J. Stoddart

Publication year - 2019

Publication title -

biotechniques

Language(s) - English

Resource type - Journals

SCImago Journal Rank - 0.617

H-Index - 131

eISSN - 1940-9818

pISSN - 0736-6205

DOI - 10.2144/btn-2019-0038

Subject(s) - trypsinization , rna , mesenchymal stem cell , biology , microrna , long non coding rna , microbiology and biotechnology , gene expression , rna extraction , non coding rna , lysis , stromal cell , gene , computational biology , genetics , biochemistry , cancer research , enzyme , trypsin

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.

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