Abstract
BackgroundDifferent methods have been used in order to isolate dental pulp stem cells. The aim of this study was to study the effect of different types of pulp treatment during isolation, under 3% O2 conditions, in the time needed and the efficacy for obtaining dental pulp stem cells.Material and MethodsOne hundred and twenty dental pulps were used to isolate dental pulp stem cells treating the pulp tissue during isolation using 9 different methods, using digestive, disgregation, or mechanical agents, or combining them. The cells were positive for CD133, Oct4, Nestin, Stro-1, CD34 markers, and negative for the hematopoietic cell marker CD-45, thus confirming the presence of mesenchymal stem cells. The efficacy of dental pulp stem cells obtention and the minimum time needed to obtain such cells comparing the 9 different methods was analyzed.ResultsDental pulp stem cells were obtained from 97 of the 120 pulps used in the study, i.e. 80.8% of the cases. They were obtained with all the methods used except with mechanical fragmentation of the pulp, where no enzymatic digestion was performed. The minimum time needed to isolate dental pulp stem cells was 8 hours, digesting with 2mg/ml EDTA for 10 minutes, 4mg/ml of type I collagenase, 4mg/ml of type II dispase for 40 minutes, 13ng/ml of thermolysine for 40 minutes and sonicating the culture for one minute.ConclusionsDental pulp stem cells were obtained in 97 cases from a series of 120 pulps. The time for obtaining dental pulp stem cells was reduced maximally, without compromising the obtention of the cells, by combining digestive, disgregation, and mechanical agents. Key words:Dental pulp stem cells, mesenchymal stem cells, isolation method.
Highlights
Dental pulp stem cells (DPSC) were first isolated by Gronthos et al [1] in 2000
Few studies analyze the influence of pulp treatment during isolation when obtaining DPSC
Our aim was to determine the time needed and the efficacy in obtaining DPSC applying 9 different pulp treatments to 120 pulps while they were being isolated under 3% O2 conditions
Summary
Dental pulp stem cells (DPSC) were first isolated by Gronthos et al [1] in 2000. These cells demonstrated clonologenic capacity and were able to differentiate into odontoblasts and form a dentin-pulp complex when they were implanted subcutaneously in immune-compromised rats. They obtained DPSC with both methodologies; cells were obtained in 3 to 5 days by enzymatic digestion of the pulp and on the fifth day with mechanical fragmentation [6] They observed no differences in terms of proliferation, colony formation or differentiation between the two methods [7]. The aim of this study was to study the effect of different types of pulp treatment during isolation, under 3% O2 conditions, in the time needed and the efficacy for obtaining dental pulp stem cells. The efficacy of dental pulp stem cells obtention and the minimum time needed to obtain such cells comparing the 9 different methods was analyzed. Results: Dental pulp stem cells were obtained from 97 of the 120 pulps used in the study, i.e. 80.8% of the cases They were obtained with all the methods used except with mechanical fragmentation of the pulp, where no enzymatic digestion was performed. The time for obtaining dental pulp stem cells was reduced maximally, without compromising the obtention of the cells, by combining digestive, disgregation, and mechanical agents
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