Abstract
This chapter describes a simplified method that allows the systematic isolation of multiple types of dental stem cells such as dental pulp stem cells (DPSC), periodontal ligament stem cells (PDLSC), and stem cells of the apical papilla (SCAP) from a single tooth. Of specific interest is the modified laboratory approach to harvest/retrieve the dental pulp tissue by minimizing trauma to DPSC by continuous irrigation, reduction of frictional heat from the bur rotation, and reduction of the bur contact time with the dentin. Also, the use of a chisel and a mallet will maximize the number of live DPSC for culture. Steps demonstrating the potential for multiple cell differentiation lineages of each type of dental stem cell into either osteocytes, adipocytes, or chondrocytes are described. Flow cytometry, with a detailed strategy for cell gating and analysis, is described to verify characteristic markers of human mesenchymal multipotent stromal cells (MSC) from DPSC, PDLSC, or SCAP for subsequent experiments in cell therapy and in tissue engineering. Overall, this method can be adapted to any laboratory with a general setup for cell culture experiments.
Key words
- Dental Pulp Stem Cells (DPSC)
- Stem Cells from Apical Papilla (SCAP)
- Periodontal Ligament Stem Cells (PDLSC)
- Mesenchymal Stromal Cells (MSC)
- Lineage differentiation
- Flow cytometry
- Cell differentiation
- Dental pulp
- Periodontal ligament
- Periapical tissue
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Acknowledgments
The authors are grateful to Xin Ran Liu for the schematic illustrations. The authors thank the Flow Cytometry and Cell Sorting facility, Department of Microbiology and Immunology, McGill University. The authors would like to thank the following funding agency: Natural Sciences and Engineering Research Council of Canada (NSERC).
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Bakkar, M. et al. (2017). A Simplified and Systematic Method to Isolate, Culture, and Characterize Multiple Types of Human Dental Stem Cells from a Single Tooth. In: Di Nardo, P., Dhingra, S., Singla, D. (eds) Adult Stem Cells. Methods in Molecular Biology, vol 1553. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-6756-8_15
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DOI: https://doi.org/10.1007/978-1-4939-6756-8_15
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