title: Influence of different types of pulp treatment during isolation in the obtention of human dental pulp stem cells.
abstract: BACKGROUND
  Different 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.
  
  
  METHODS
  One 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.
  
  
  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 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.
  
  
  CONCLUSIONS
  Dental 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.
authors.affiliation: Cirugía Bucal, Clínicas Odontológicas, Facultad de Medicina y Odontología, Universidad de Valencia, Gascó Oliag 1, E46021- Valencia, Spain, miguel.penarrocha@uv.es.
authors.forename: J
authors.initials: J
authors.lastname: Viña-Almunia
authors.forename: C
authors.initials: C
authors.lastname: Borras
authors.forename: J
authors.initials: J
authors.lastname: Gambini
authors.forename: M
authors.initials: M
authors.lastname: El Alamy
authors.forename: M
authors.initials: M
authors.lastname: Peñarrocha
authors.forename: J
authors.initials: J
authors.lastname: Viña
mesh_terms: D002478:Cells, Cultured; D003782:Dental Pulp; D004847:Epithelial Cells; D006801:Humans; D059630:Mesenchymal Stem Cells; D033581:Stem Cell Transplantation