Isolation and Characterization of hIDPSCs

The hIDPSCs used in this study were isolated from the dental pulp of deciduous teeth of a 6-year-old individual (male), using a novel and disruptive technology developed by Kerkis et al. [18 (link)] and protected by the US patent number US20160184366A1. The hIDPSCs were cultivated until the fifth passage in basal medium (Dulbecco’s modified Eagle’s medium (DMEM)/Ham’s F12, supplemented with 15% fetal bovine serum, 100 U/mL penicillin, 100 μg/mL streptomycin, 2 mM L-glutamine and 2 nM nonessential amino acids, all from Gibco, Carlsbad, CA, USA) [23 (link)]. The MSC phenotype of these cells was confirmed using the criteria defined by the International Society for Cellular Therapy (ISCT) [24 (link),25 (link)]. The hIDPSCs employed in this study are CD105-, CD73- and CD90-positive, and CD45-, CD34-, CD11b- and HLA-DR-negative, as previously described by Kerkis et al. [18 (link)]. These cells also express high levels of BDNF, as previously demonstrated [26 (link)]. The hIDPSCs employed in this study were produced by the Brazilian facility Cellavita Pesquisas Científicas Ltda. (Valinhos-SP, Brazil) according to the good manufacturing practices (GMP) required by the Brazilian Health Regulatory Agency (ANVISA, RDC 508/21) for advanced therapy products [23 (link)]. These cells comprise the active component of the NestaCell® product [23 (link)].