Dimethyl sulfoxide (dmso)

Before cryopreservation, DPSCs/ADSCs of each lineage were divided into seven aliquots containing 0.5 × 10⁶ cells according to the CPA/s supplemented in the cryopreservation media (Table 1).
The cryovials containing a pharmacological grade high-molecular-weight HA were distributed and preprepared by the company Contipro a.s., Dolni Dobrouc, Czech Republic (Figure 1).
First, we filled the experiment cryovials containing the HMW-HA with 0.5 mL of the modified cultivation media for mesenchymal adult progenitor cells minus the volume of cell aliquots containing 0.5 × 10⁶ cells submerged in the modified cultivation media. This was undertaken to dissolve any HMW-HA present before we placed the cells in each cryovial. Subsequently, we added the cell samples and 0.5 mL of precooled 4 °C cryopreservation medium composed of DMSO (Sigma-Aldrich—Merck KGaA) and FBS (PAA Laboratories) according to final concentrations of DMSO in the experiment groups, namely 5%, and 3%. In the control groups, we followed the same cryopreservation protocols but without the HMW-HA.
Afterward, we stored each cryovial using uncontrolled-rate freezing. The cryovials were placed at a temperature of −20 °C and kept for 1–1.5 h. Then, they were placed directly in the freezer and stored at −80 °C for one week.

The chemical composition of PAH-derived SOA is quite complex, with hundreds to thousands of compounds being present. To simplify our approach, and to allow a molecular approach to the toxic response of these particles, we investigated the impact of by-products mimicking naphthalene-derived aerosols [7 (link),8 (link),9 (link),10 (link),30 (link)].
The chemicals investigated in this study were 1,4-naphthoquinone (1,4-NQ), 2-hydroxy-1,4 naphthoquinone (2-OH-NQ), phthalic acid (PA) and phthaldialdehyde (OPA), purchased from Sigma-Aldrich (St. Quentin Fallavier, France). All chemicals were dissolved in DMSO and further diluted in Dulbecco’s Modified Eagle’s Medium with low glucose (1 g/L) containing 10% fetal calf serum (DMEM 10% FCS, PAA Laboratories, Toronto, ON, Canada) with the addition of streptomycin plus penicillin (100 units/mL; Sigma Aldrich, St-Louis, MO, USA) with 0.1% of DMSO (Sigma Aldrich, St-Louis, MO,USA) in the end.
Cells were exposed to different concentrations of the compounds when the cell index reached the value of 1.0. The concentrations tested for 1,4-NQ were 12.5, 25, 50 and 100 µM. The concentrations for 2-OH-NQ, phthalic acid (PA) and phthaldialdehyde (OPA) were 1, 2.5, 5, 7.5 and 10 mM.

Aβ_(1–42) peptide (DAEFRHDSGYEVHHQKLVFFAEDVGSNKGAIIGLMVGGVVIA) or Aβ_(1–42) scramble peptide (KVKGLIDGAHIGDLVYEFMDSNSAIFREGVGAGHVHVAQVEF; rPeptide, Bogart, Georgia, USA) solution were prepared as described previously (Kuperstein et al., 2010 (link); Broersen et al., 2011 (link); Brouillette et al., 2012 (link)). Vials containing 0.5 mg of Aβo_(1–42)–HFIP (1, 1, 1, 3, 3, 3-Hexafluoro-2-propanol) films were allowed to defrost at room temperature for 10 min. Aβo_(1–42) was then dissolved in HFIP (Sigma-Aldrich) at a concentration of 1 mg/mL. The HFIP was evaporated using a gentle steam of nitrogen gas and the peptide film was dissolved in 500 μL of DMSO (Sigma-Aldrich). DMSO was totally removed of the solution by eluting the Aβo_(1–42) peptide from a 5 ml HiTrap desalting column (GE Healthcare) with 50 mL of buffer (pH 7.5) containing (Tris50 mM and EDTA 1 mM). Aβo_(1–42) peptide was then kept at −80°C in 30 μL aliquots until use. The concentration of Aβo_(1–42)in each sample was measured using a BCA protein assay kit (Pierce, USA).