Bioanalyzer equipment

To compare the phenotype of the undifferentiated cells, RNA extraction was performed from DPSCs (n = 6) and ADSCs (n = 6), between passages three and six, according to the protocol described in a commercial kit (PureLink RNA Mini Kit—Life Technologies, Carlsbad, CA, USA). Cell lysis, purification and RNA elution were performed.
Samples were treated with DNase (RNase-Free DNase Set- Qiagen, Germantown, MD, USA). The RNA was quantified in a spectrophotometer (Nanodrop-Thermo Scientific, Wilmington, DE, USA), and the quality was analysed with Bioanalyzer equipment (Agilent, Santa Clara, CA, USA). Only samples with an RNA integrity number (RIN) better than 7 were considered for sequencing.
Samples were sent to the Edinburgh Genomics sequencing facility (Edinburgh, UK) and multiplexed and sequenced on Illumina HiSeq 4000 equipment (Illumina Inc., San Diego, CA, USA) to generate paired-end fragments of 100 base pairs.

Cells were detached with Tryple Express (Invitrogen, Carlsbad, CA, USA), washed twice with PBS and their RNA was isolated using TriZOL LS Reagent, according to manufacturer’s instructions. The RNAs used for microarrays were checked for quality in the Bioanalyzer equipment (Agilent Technologies). Only RNAs with RIN (RNA Integrity Number) values greater than 7 were used for microarrays.