Isolation and Characterization of Dental Pulp Stem Cells

Dental pulp was harvested from the extracted incisors of 6-week-old male green fluorescent protein (GFP)-transgenic SD rats (SD-Tg(CAG-EGFP)Cz-0040sb) (Japan SLC, Inc., Hamamatsu, Japan) and suspended in phosphate-buffered saline containing 0.1% collagenase and 0.25% trypsin- ethylenediaminetetraacetic acid (EDTA). DPSCs were cultured in an alpha modification of Eagle’s medium (α-MEM) (GIBCO, Thermo Fisher Scientific, Waltham, MA, USA) with 20% fetal bovine serum (GIBCO) on plastic dishes at 37 °C in 5% humidified CO₂. Non-adherent cells were washed off, and adherent cells were continuously expanded until the third passage. DPSCs expressed the cell surface markers of mesenchymal stem cells, such as CD29 and CD90, and their pluripotency to chondrocytes, adipocytes, and osteoblasts was confirmed [13 (link)].