Mscgm cd medium

Cells derived from periodontal tissue were collected as previously described, scraping a third coronal root surface using Gracey’s curette^{33 (link)}. All three patients, included in the study, were in good general health and exempt from oral and systemic diseases. After collection, hPDLSCs were cultured using MSCGM-CD medium (mesenchymal stem cell growth medium chemically defined) (Lonza, Basel, Switzerland) and were maintained in an incubator at 37 °C in a humidified atmosphere of 5% CO2 in air. To evaluate cells morphological features, plastic-adherent hPDLSCs were stained used toluidine blue solution and then observed at inverted light microscopy DMIL (Leica Microsystem, Milan, Italy)³⁴ .

hPDLSCs were isolated from the periodontal tissue and hDPSCs from the dental pulp of noncarious third molars extracted for orthodontic purpose, as previously described [19 (link)]. Gingival tissues were collected during surgical gingival resection for the extraction of a supernumerary tooth or for orthodontic reasons. hGMSCs were detached after their spontaneous migration from tissue samples as reported by Soundara Rajan et al. [20 (link)]. All donors were in good general health and exempt from oral and systemic diseases. Cells were cultured using MSCGM-CD medium (mesenchymal stem cell growth medium chemically defined) (Lonza, Basel, Switzerland) and were maintained in an incubator at 37°C in a humidified atmosphere of 5% CO₂ in air. Cells were subcultured until P2 and P15. All experiments were performed in triplicate. Cells derived from each donor have been used separately.

The primary human DPSCs used in this study were isolated in the Dental Clinics of the Medical School of University “G. d’Annunzio” Chieti-Pescara. Human DPSCs were collected from a third molar scheduled to be extracted for orthodontic treatment as previously reported [17 (link)]. Tissue fragments were cultured for one week in a humidified and controlled atmosphere of 37 °C and 5% CO₂ with MSCGM-CD medium (mesenchymal stem-cell growth medium chemically defined) (Lonza, Basel, Switzerland). Cells spontaneously migrated from tissue fragments were subcultured until P2. The medium was refreshed every two days. The cells used in all the experiments were at passage 2. All experiments were performed in triplicate.