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Analysis of side population cells derived from dental pulp tissue
Author(s) -
Kenmotsu M.,
Matsuzaka K.,
Kokubu E.,
Azuma T.,
Inoue T.
Publication year - 2010
Publication title -
international endodontic journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.988
H-Index - 119
eISSN - 1365-2591
pISSN - 0143-2885
DOI - 10.1111/j.1365-2591.2010.01789.x
Subject(s) - nestin , side population , pulp (tooth) , collagenase , alkaline phosphatase , population , cell counting , andrology , cell , dispase , biology , chemistry , microbiology and biotechnology , stem cell , pathology , cell cycle , medicine , biochemistry , neural stem cell , environmental health , cancer stem cell , enzyme
Kenmotsu M, Matsuzaka K, Kokubu E, Azuma T, Inoue T. Analysis of side population cells derived from dental pulp tissue. International Endodontic Journal . Abstract Aim To investigate the characteristics of side population (SP) cells derived from the dental pulp of young and aged rats. Methodology Maxillary and mandibular incisors were extracted from 5‐week‐old (young) rats and 60‐ to 80‐week‐old (aged) rats. Coronal pulp tissue was removed mechanically, and single‐cell suspensions were prepared using collagenase and dispase. Cells were stained with Hoechst 33342 and sorted with an fluorescence‐activated cell sorter (FACS). Isolated SP and main population (MP) cells were analysed by real‐time reverse transcription polymerase chain reaction, immunohistochemical localization and cell cycle determination. Two‐way analysis of variance and the multiple comparison Scheffè test were used for statistical analysis ( P < 0.05). Results Approximately 0.40% of pulp cells in young rats and 0.11% in aged rats comprised SP cells. SP cells expressed a higher mRNA level of ATP‐binding cassette transporter G2 (ABCG2), but lower mRNA levels of nestin, alkaline phosphatase, p16 and p57 than MP cells in both age groups. Immunohistochemical observation revealed ABCG2‐positive cells localized in the cell‐rich zone and nestin in the odontoblastic layer in both groups. Furthermore, the majority of both young and aged SP and MP cells were in growth arrest of the G 0 /G 1 phase. Conclusion The FACS analysis revealed a decrease in the proportion of SP cells with age, whilst p16 mRNA expression indicated an increase in cell senescence. The cell cycles of SP and MP cells from both young and aged dental pulp were generally in the G0/G1 phase.