Proliferation rate and expression of stem cells markers during expansion in primary culture of pulp cells

Autor(es)

Turrioni, Ana Paula ^[1] ; de Oliveira Neto, Nilson Ferreira ^[1] ; Xu, Yan ^[2] ; Morse, Leslie ^[3] ; de Souza Costa, Carlos Alberto ^[4] ; Battaglino, Ricardo ^[3] ; Hebling, Josimeri ^[5]

Afiliação do(s) autor(es)

^[1] Univ Fed Uberlandia UFU, Sch Dent, Dept Pediat Dent, Uberlandia, MG - Brazil

^[2] Forsyth Inst, Dept Mineralized Tissue Biol, Cambridge, MA - USA

^[3] Univ Minnesota, Sch Med, Dept Rehabil Med, Minneapolis, MI - USA

^[4] Univ Estadual Paulista, UNESP, Sch Dent, Dept Physiol & Pathol, Araraquara, SP - Brazil

^[5] Univ Estadual Paulista, UNESP, Sch Dent, Dept Orthodont & Pediat Dent, Araraquara, SP - Brazil

• Tipo de documento: Artigo Científico
• Fonte: Brazilian Oral Research; v. 35, 2021.
• Citações Web of Science: 0

Resumo

Abstract: The aim of the present study was to evaluate the proliferation rate and the expression of stem cells markers during expansion in primary culture of dental pulp stem cells (DPSCs), comparing different techniques (explant and enzymatic digestion), subject ages (up to 40 and over 40) and cell passages (#2, #5 and #8). DPSCs were isolated using either the enzymatic digestion (ED) or explant (EX) technique. The number of days needed for the cells to reach confluence was determined. Immunophenotyping was performed by immunofluorescence and flow cytometry analysis using antibodies specific for nestin, vimentin, CD44, CD146, Oct3/4 and CD34. Data were subjected to three-way analysis of variance (n = 6/group). The ANOVA tests were complemented by Tukey's or t-tests (p < 0.05). The variables “donor age” and “technique” were analyzed to define the optimal desirability value using a response optimization. DPSCs presented a high proliferation rate from passages 2 to 5 while cells from passage 8 proliferated at a slower rate. For all markers, no significant difference was observed among passages, irrespective of the technique used or the donor's age. The mean fraction of specific antibodies was 73.7% (± 11.5), 49.0% (± 18.7), 80.1% (± 8.0), 45.2% (± 13.7), 64.7% (± 5.3) and 2.0% (± 1.5) for CD44, OCT, vimentin, nestin, CD146 and CD34, respectively. The highest optimal desirability value was obtained using the ED technique and cells from younger patients (d = 0.92). However, it was concluded that neither the isolation technique nor the donor age or cell passage significantly interfered with the stem cell phenotype and proliferation rate during cell expansion. (AU)

• Processo FAPESP: 13/17758-3 - Efeito da técnica de isolamento, fonte do tecido e passagem celular no número e fenótipo de células-tronco originárias da polpa dentária
• Beneficiário: Ana Paula Silveira Turrioni
• Modalidade de apoio: Bolsas no Exterior - Estágio de Pesquisa - Doutorado