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Cell Journal، جلد ۱۵، شماره Suppl ۱، صفحات ۳۱-۳۱
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عنوان انگلیسی Ps-26: Optimized Method for Isolation of Stem Cells from Human Dental Pulp
چکیده انگلیسی مقاله Objective: Dental pulp stem cells (DPSCs) are a population of clonogenic and highly proliferative cells derived from enzymatically digested Dental pulp tissue regularly. Providing scaling-up of stem cells at early passages is of importance for regenerative medicine purposes. Typically, two protocols are employed to isolate stem cells from human dental pulp: tissue explants culture and culture of released cells from enzymatically digested pulp tissue. The present study compared two major isolation methods and enhanced efficiency of cell isolation by some modifications. Materials and Methods: Dental pulp was extracted from third molars of 60 healthy subjects. In the first method pulp digested with 1 and 3 mg/mL collagenase/dispase (Roche) for 30 and 60 minutes at 37 °C and released cells obtained using a 70-μm cell strainer for culture, in the second method intact pieces of pulp cultured. In the third method digested pulp pieces from 2 enzyme concentrations for 30 and 60 minutes were immobilized and cultured. The cells and tissues maintained in alpha-MEM supplemented with 20% fetal bovine serum (FBS), 100 U/mL penicillin, 100μg/mL streptomycin, and 25 ng/mL amphotericin B and incubated in humidified incubator with 5% CO2 at 37 °C. The outgrowing and cultured cells monitored visually. In each group cells and colonies were counted and compared. Results: Fiftheen different modifications in isolation methods were examined. The results showed that treating pulp segments with1 mg/mL collagenase/dispase for 30 minutes and culturing immobilized tissues increased the efficiency of cell isolation up to 80% and cells appear in 3-4 days of culture compared with other methods < 20% in 10-15 days. Conclusion: According to the small size of pulp tissue and its low stem cell contents, acquiring substantial quantities of cells in primary culture will facilitate the in vitro expansion and providing adequate production of the stem cells at early passages with minimum risk of losing their ‘‘stemness’’ and aberrant genetic changes for use in research, tissue engineering and regenerative medicine. Optimized method increase efficiency of cell isolation and provides significant quantities of stem cells in primary culture more than other methods.
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نشانی اینترنتی http://celljournal.org/journal/article/abstract/831
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