Isolation of Human Periodontal Ligament Stem Cells

To collect hPDLSCs, we have enrolled six patients in good general health and without oral cavity diseases. They were to undergo surgical procedures to start the orthodontic treatment. After the collection, the periodontal ligament fragments were washed five times with phosphate-buffered saline solution (PBS, Lonza, Basel, Switzerland) [28 (link)]. The washed tissue fragments were placed in a culture dish in an incubator at 37°C in a humidified atmosphere of 5% CO₂ in air. The mesenchymal stem cell growth medium-chemically defined (MSCGM-CD, Lonza) was used as a medium and changed every two days. Isolated cells were migrated spontaneously from the tissue fragments after two weeks of culture [29 (link), 30 (link)].