title: A Simplified and Systematic Method to Isolate, Culture, and Characterize Multiple Types of Human Dental Stem Cells from a Single Tooth.
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.
authors.affiliation: Craniofacial Tissue Engineering and Stem Cells Laboratory, Faculty of Dentistry, McGill University, 3640 University Street, M43, Montreal, QC, Canada.
authors.forename: Mohammed
authors.initials: M
authors.lastname: Bakkar
authors.affiliation: Craniofacial Tissue Engineering and Stem Cells Laboratory, Faculty of Dentistry, McGill University, 3640 University Street, M43, Montreal, QC, Canada.
authors.forename: Younan
authors.initials: Y
authors.lastname: Liu
authors.affiliation: Craniofacial Tissue Engineering and Stem Cells Laboratory, Faculty of Dentistry, McGill University, 3640 University Street, M43, Montreal, QC, Canada.
authors.forename: Dongdong
authors.initials: D
authors.lastname: Fang
authors.affiliation: Craniofacial Tissue Engineering and Stem Cells Laboratory, Faculty of Dentistry, McGill University, 3640 University Street, M43, Montreal, QC, Canada.
authors.forename: Camille
authors.initials: C
authors.lastname: Stegen
authors.affiliation: Craniofacial Tissue Engineering and Stem Cells Laboratory, Faculty of Dentistry, McGill University, 3640 University Street, M43, Montreal, QC, Canada.
authors.forename: Xinyun
authors.initials: X
authors.lastname: Su
authors.affiliation: Craniofacial Tissue Engineering and Stem Cells Laboratory, Faculty of Dentistry, McGill University, 3640 University Street, M43, Montreal, QC, Canada.
authors.forename: Murali
authors.initials: M
authors.lastname: Ramamoorthi
authors.affiliation: Craniofacial Tissue Engineering and Stem Cells Laboratory, Faculty of Dentistry, McGill University, 3640 University Street, M43, Montreal, QC, Canada.
authors.forename: Li-Chieh
authors.initials: LC
authors.lastname: Lin
authors.affiliation: Department of Regenerative Oral Surgery, Nagasaki University Graduate School of Biomedical Sciences, Nagasaki, Japan.
authors.forename: Takako
authors.initials: T
authors.lastname: Kawasaki
authors.affiliation: Craniofacial Tissue Engineering and Stem Cells Laboratory, Faculty of Dentistry, McGill University, 3640 University Street, M43, Montreal, QC, Canada.
authors.forename: Nicholas
authors.initials: N
authors.lastname: Makhoul
authors.affiliation: Craniofacial Tissue Engineering and Stem Cells Laboratory, Faculty of Dentistry, McGill University, 3640 University Street, M43, Montreal, QC, Canada.
authors.forename: Huan
authors.initials: H
authors.lastname: Pham
authors.affiliation: Department of Regenerative Oral Surgery, Nagasaki University Graduate School of Biomedical Sciences, Nagasaki, Japan.
authors.forename: Yoshinori
authors.initials: Y
authors.lastname: Sumita
authors.affiliation: Craniofacial Tissue Engineering and Stem Cells Laboratory, Faculty of Dentistry, McGill University, 3640 University Street, M43, Montreal, QC, Canada. simon.tran@mcgill.ca.
authors.forename: Simon D
authors.initials: SD
authors.lastname: Tran
mesh_terms: D015415:Biomarkers; D018929:Cell Culture Techniques; D002454:Cell Differentiation; D019070:Cell Lineage; D049109:Cell Proliferation; D002469:Cell Separation / Q000379:methods; D015925:Cryopreservation / Q000379:methods; D003782:Dental Pulp / Q000166:cytology; D006801:Humans; D016130:Immunophenotyping; D059630:Mesenchymal Stem Cells / Q000166:cytology* / Q000378:metabolism*; D010513:Periodontal Ligament / Q000166:cytology; D010641:Phenotype; D014070:Tooth / Q000166:cytology; D057188:Workflow
chemical_list: D015415:Biomarkers
keywords: Cell differentiation; Dental Pulp Stem Cells (DPSC); Dental pulp; Flow cytometry; Lineage differentiation; Mesenchymal Stromal Cells (MSC); Periapical tissue; Periodontal Ligament Stem Cells (PDLSC); Periodontal ligament; Stem Cells from Apical Papilla (SCAP)