Characterization of Immune-Selected hDPSCs

In order to confirm the phenotype of the immune-selected hDPSCs the expression of the stemness markers STRO-1 and c-Kit, the neural crest-related antigens nestin and SOX10, and the immunomodulatory molecule FasL was investigated. Subsequently, the multipotency of the STRO-1⁺/c-Kit⁺ hDPSCs population was demonstrated by culturing cells in appropriate differentiation media to reach osteogenic, myogenic and glial differentiation, respectively, as formerly described (Di Tinco et al., 2021a; Zordani et al., 2019 (link); Carnevale et al., 2018 (link)). At the end of each differentiation experiment, the commitment was evaluated by assessing the expression of lineage related markers with the use of the following primary antibodies: mouse anti-osteocalcin (OCN), rabbit anti-RUNX2 (Abcam), mouse anti-myogenin, rabbit anti-desmin and rabbit anti-S100b (all from Sigma Aldrich). Confocal immunofluorescence analyses were performed as detailed below.

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Pisciotta A., Lunghi A., Bertani G., Di Tinco R., Bertoni L., Orlandi G., Biscarini F., Bianchi M, & Carnevale G. (2022). PEDOT: PSS promotes neurogenic commitment of neural crest-derived stem cells. Frontiers in Physiology, 13, 930804.

Publication 2022

mouse anti-myogenin rabbit anti-desmin rabbit anti-S100b

Antibodies Antigens C kit Desmin Fasl Glial Immunofluorescence Immunomodulatory Mouse Myogenic Myogenin Nestin Neural crest Osteocalcin Osteogenic Phenotype Rabbit Runx2 Sox10

Corresponding Organization : Italian Institute of Technology

Other organizations : University of Modena and Reggio Emilia, Istituto Nazionale di Fisica Nucleare, Sezione di Ferrara

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Variable analysis

independent variables

Culture of hDPSCs in appropriate differentiation media to reach osteogenic, myogenic and glial differentiation

dependent variables

Expression of lineage-related markers (osteocalcin, RUNX2, myogenin, desmin, S100b)

control variables

Experimental procedures as formerly described in the cited references (Di Tinco et al., 2021a; Zordani et al., 2019; Carnevale et al., 2018)

controls

Positive controls: Not explicitly mentioned
Negative controls: Not explicitly mentioned

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