Cell Journal (Jan 2008)
Isolation of Mesenchymal Stem Cells From Dental Pulp of Exfoliated Human Deciduous Teeth
Abstract
Objective: The exfoliated human deciduous tooth (SHED) contain multipotent stemcells that identified to be a population of highly proliferative and clonogenic .Thesecells are capable of differentiating into a variety of cell types including neural cells,adipocytes, and odontoblasts.Material and Methods: Normal exfoliated human deciduous incisors collected fromsix- to nine-years-old children. The pulp was separated from the crown and digestedwith collagenase .Single cell solutions were cultivated in α-MEM supplemented withES-FCS. After two to three days, the cells reached confluency and were trypsinizedand cultured for further passages. The passage-4 cells were analyzed with CD34,CD45, CD105, CD166, CD31, CD90 and CD146 markers that indicated these cellshad a mesenchymal stem cell (MSC) identity. We examined the cells for AlkalinePhosphatase activity to investigate the mesenchymal (stromal) nature.Finally, thecells were differentiated into the osteoblastic and adipocytic lineages in differentsubcultures and analysed by RT-PCR and different staining protocols.Results: Viable cells growing out of the explants showed elongated shapes inclusters. These cells showed alkaline phosphatase activity. Flow cytometry resultsrevealed high expression of pluripotent stem cell markers .In some area of theosteoinductive cultures nodule-like structures were observed that showed redmineralizing area upon staining with Alizarin Red.In adipogenic cultures lipid vesiclesappeared after five weeks of induction with Oil Red.Conclusion: This study show that pulp contains cells with high plasticity andproliferation capacity and can be easily isolated without any serious intervention.
