Isolation and Characterization of Dental Pulp Stem Cells from Murine Incisors

Akmal, Mohammad; Zarina, Z. A. Intan; Rohaya, M. A. W.; Sahidan, S.; Ariffin, Shahrul Hisham Zainal; Hisham, Z. A. Shahrul

doi:10.3923/jbs.2014.327.331

Cited by 6 publications

(7 citation statements)

References 14 publications

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“…Outgrowth and enzymatic digestion methods are two commonly used methods to obtain dental pulp stem cells (DPSC). Enzymatic digestion allows the release of all types of cells from the pulp tissue, resulting in cells with different morphologies to exist at passage 0 ( Nur Akmal et al, 2014 ; Yildirim, 2013b ). While more homogenous cell morphology can be obtained through the outgrowth method, there is the possibility of spontaneous differentiation taking place as the cells that migrate out might grow densely around the tissue.…”

Section: Discussionmentioning

confidence: 99%

“…Stem cells are able to spontaneously differentiate and expresses mature cell markers when maintained in confluent state ( Dudakovic et al, 2014 ). As these cells may consist of both mature and stem cells, passaging of cells makes it possible for highly proliferative cells, and cells that can continuously divide, to be obtained ( Nur Akmal et al, 2014 ; Yildirim, 2013b ). The fibroblastic morphology obtained is a common dental pulp stem cell morphology as previously reported ( Yildirim, 2013b ).…”

Section: Discussionmentioning

confidence: 99%

“…RT-PCR was conducted by means of touchdown PCR using a Techne® Prime thermal cycler using the primers and steps previously published in detail (Nur Akmal et al, 2014). …”

Section: Methodsmentioning

confidence: 99%

“…Total RNA was isolated and converted to cDNA using a Grandscript cDNA Synthesis Kit (PCR Biosystems, UK). RT-PCR was conducted by means of touchdown PCR using a Techne® Prime thermal cycler using the primers and steps previously published in detail ( Nur Akmal et al, 2014 ).…”

Section: Methodsmentioning

confidence: 99%

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Impact of isolation method on doubling time and the quality of chondrocyte and osteoblast differentiated from murine dental pulp stem cells

Wahab

Rozali

Senafi

et al. 2017

PeerJ

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BackgroundStem cells are normally isolated from dental pulps using the enzymatic digestion or the outgrowth method. However, the effects of the isolation method on the quality of the isolated stem cells are not studied in detail in murine models. The aim of this study was to compare the matrices secreted by osteoblast and chondrocytes differentiated from dental pulp stem cells isolated through different means.MethodDPSC from murine incisors were isolated through either the outgrowth (DPSC-OG) or the enzymatic digestion (DPSC-ED) method. Cells at passage 4 were used in this study. The cells were characterized through morphology and expression of cell surface markers. The cells’ doubling time when cultured using different seeding densities was calculated and analyzed using one-way ANOVA and Tukey’s multiple comparison post-test. The ability of cells to differentiate to chondrocyte and osteoblast was evaluated through staining and analysis on the matrices secreted.ResultsGene expression analysis showed that DPSC-OG and DPSC-ED expressed dental pulp mesenchymal stem cell markers, but not hematopoietic stem cell markers. The least number of cells that could have been used to culture DPSC-OG and DPSC-ED with the shortest doubling time was 5 × 102 cells/cm2 (11.49 ± 2.16 h) and 1 × 102 cells/cm2 (10.55 h ± 0.50), respectively. Chondrocytes differentiated from DPSC-ED produced 2 times more proteoglycan and at a faster rate than DPSC-OG. FTIR revealed that DPSC-ED differentiated into osteoblast also secreted matrix, which more resembled a calvaria.DiscussionIsolation approaches might have influenced the cell populations obtained. This, in turn, resulted in cells with different proliferation and differentiation capability. While both DPSC-OG and DPSC-ED expressed mesenchymal stem cell markers, the percentage of cells carrying each marker might have differed between the two methods. Regardless, enzymatic digestion clearly yielded cells with better characteristics than outgrowth.

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

“…RT-PCR was conducted by means of touchdown PCR using a Techne® Prime thermal cycler using the primers and steps previously published in detail (Nur Akmal et al, 2014). …”

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

See 2 more Smart Citations

Impact of isolation method on doubling time and the quality of chondrocyte and osteoblast differentiated from murine dental pulp stem cells

Wahab

Rozali

Senafi

et al. 2017

PeerJ

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“…A robust number of studies have investigated the origin, differentiation, organogenesis and regeneration of teeth. There has been a huge progress in recent years towards a much better understanding of the regulation of tooth development (20)(21)(22)(23).…”

Section: Literature Review Toothmentioning

confidence: 99%

Comparative gene expression analysis of human dental pulp cells from maxillary and mandibular teeth

Faruangsaeng¹

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Objective : Given the anatomical and locational differences between the maxillary and mandibular teeth, it is possible that there are differences in gene expression patterns of dental pulp cells within those teeth. This study evaluates and compares the gene expression profiles of the maxillary and mandibular human teeth with the aim of finding differences in gene expression patterns of dental pulp cells by RNA-Sequencing technique. Methods : The upper and lower premolar and molar teeth obtained from the same participant were employed. Two pairs of opposing premolar teeth and two pairs of opposing molar teeth were isolated, cultured, and subjected for RNA sequencing. RNA-Seq Alignment and RNA-Seq Differential Expression were used to analyze gene expression profile. Quantitative RT-PCR was performed to confirm the result obtained from RNA-Sequencing. Results : RNA sequencing demonstrated that 19,372 genes out of 27,914 in total were expressed. The top expression genes were FN1, COL1A1, COL1A2, ACTB and EEF1A1. In DPSCs of posterior teeth (premolar and molar), only PITX1 gene had a significant higher expression (exhibiting 26.47 or 89-fold change) in the lower teeth compared to the upper teeth (64-fold change in premolar teeth and 116-fold change in molar teeth). Conclusion : We showed that PITX1 gene had a significant higher expression in the lower teeth compared to the upper teeth and this difference in PITX1 level was more evident in the molars compared to premolars.

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Dual growth factor delivery using PLGA nanoparticles in silk fibroin/PEGDMA hydrogels for articular cartilage tissue engineering

et al. 2019

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Isolation and Characterization of Dental Pulp Stem Cells from Murine Incisors

Cited by 6 publications

References 14 publications

Impact of isolation method on doubling time and the quality of chondrocyte and osteoblast differentiated from murine dental pulp stem cells

Impact of isolation method on doubling time and the quality of chondrocyte and osteoblast differentiated from murine dental pulp stem cells

Comparative gene expression analysis of human dental pulp cells from maxillary and mandibular teeth

Dual growth factor delivery using PLGA nanoparticles in silk fibroin/PEGDMA hydrogels for articular cartilage tissue engineering

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