Immunocytochemical characterization of primary teeth pulp stem cells from three stages of resorption in serum‐free medium

Abstract

Background/Aims: Dental pulp stem cells from primary teeth cultured in serum-free conditions may have clinical use for the repair and regeneration of teeth as well as other complex tissues and organs. The aim of this study was to test the change in the stem cell markers expression/ stem cell population in human primary pulp cells at the different stages of root resorption. Methods: Caries-free human primary canines at defined stages of physiological root resorption were included (n = 9). In vitro cultures were established in xeno-free, serum-free Essential 8™ medium with human truncated vitronectin for cell attachment. An embryonic stem cell line (GENEA002) was used as a positive control. The expression of embryonic stem cell markers (Oct4, Nanog and Sox2), neural crest stem cell markers (nestin and Dlx2) and mesenchymal stem cell surface markers (CD90, CD73 and CD105) were investigated by immunocytochemistry. Mesenchymal stem cell markers CD105, CD73 and CD90 and haematopoietic markers: CD45, CD34, CD11b, CD19 and HLA-DR were quantified with flow cytometry. Results: The early neural progenitor markers nestin and Dlx2 were detected in most serum-free cultured dental pulp stem cells, regardless of the tooth resorption stage from which they were harvested. Only isolated cells were found that expressed the embryonic stem cell transcription factors Oct4A, Nanog and Sox2, and in the late stages of resorption, no Oct4A was detected. The majority expressed the mesenchymal stem cell markers CD90, CD73 and CD105. Flow cytometry found positive signals for CD90 > 97.3%, CD73 > 99.6% and CD105 > 82.5%, with no detectable differences between resorption stages. Conclusions: This study identified populations of dental pulp cells in vitro with markers characteristically associated with embryonic stem cells, neural crest-derived cells and mesenchymal stem cells. Flow cytometry found CD105 expressed at lower levels than CD90 and CD73. The consistency of stem cell marker expression in cells cultured from teeth at different resorption stages suggests that pre-exfoliated primary teeth that are free of caries may provide a convenient source of multipotent stem cells for use in regenerative medicine.

Comparative Analysis of Dental Pulp and Periodontal Stem Cells: Differences in Morphology, Functionality, Osteogenic Differentiation and Proteome

“…DPSC are known to derive from neural crest cells and are inclined to differentiate into neural cells [ 36 ]. DPSC have a higher positive ratio for neural markers such as NES, GFAP, and s100-beta than other kinds of MSC [ 5 , 36 , 70 , 71 ]. However, PDLSC have different embryonic origin: dental pulp is formed from dental papilla while PDLSC originate from dental follicle cells [ 7 , 72 ].…”

Mesenchymal stromal cells derived from exfoliated deciduous teeth express neuronal markers before differentiation induction

“…SHED isolation efficiency was 100% and we observed no contamination during cell cultivation. Alansary, et al 19 (2020) collected teeth with different levels of root resorption and observed contamination in the culture of half of the samples, even using antibiotics and antimycotics. The authors claimed that teeth with advanced root resorption have a higher chance of contamination during cultivation.…”

“… 2 Studies observed a lower expression of CD105 in SHED, an important marker of the panel suggested by the ISCT. 2 , 19 – 21 This low expression is possibly due to the time of teeth collection (stages of root resorption) and the different isolation and culture methods. According to Alansany, et al 19 (2020), the lowest expression of CD105 occurs in cells grown in serum-free conditions.…”

“… 2 , 19 – 21 This low expression is possibly due to the time of teeth collection (stages of root resorption) and the different isolation and culture methods. According to Alansany, et al 19 (2020), the lowest expression of CD105 occurs in cells grown in serum-free conditions. However, based on so many variables, more studies on CD105 must be carried out to understand better which variable acts in the expression.…”

Relationship between Physiological Resorption of Primary Molars with Its Permanent Successors, Dental Age and Chronological Age

“…Rowlands et al also state that premolar eruption can be predicted from CRR [ 48 ]. The current lines of research analyse the morphological changes of the pulp cells [ 49 ] and ultrastructural alterations in the pulp tissues [ 50 ] during the different stages of physiological root resorption, which will help to better understand this process.…”