Isolation and characterization of human dental pulp stem cells (DPSCs) and stem cells from human exfoliated deciduous teeth (SHED) – an in vitro study

2018 ◽  
Vol 10 (2) ◽  
pp. 9-18
Author(s):  
Dr. Vinay Rao ◽  
Dr. Roopa R Nadig ◽  
Dr.Ramanand Nadig ◽  
Dr.Karthik .J ◽  
Dr. Veena Pai S
Keyword(s):  
Stem Cells ◽  
Dental Pulp ◽  
Surface Characterization ◽  
Dental Pulp Stem Cells ◽  
In Vivo Studies ◽  
Deciduous Teeth ◽  
Pulp Tissue ◽  
Isolation And Characterization

AIMS AND OBJECTIVES: 1. Isolation and growth of dental pulp stem cells (DPSCs) and stem cells from exfoliated human deciduous teeth (SHED). 2. Characterization of dental pulp stem cells (DPSCs) and stem cells from exfoliated human deciduous teeth (SHED). METHODS: The pulp tissue was digested in collagenase and cultured in DMEM Dulbecco’s Modified Eagle’s Media). The stem cells were identified and isolated. Surface characterization of cells was done with the help of flow cytometer using a panel of various surface markers. An immuno cytochemistry analysis was done to see the expression of proteins in the cells. RESULTS: Identification of cells was done with the help of a phase contrast microscope. Flow cytometry analyses for various CD markers showed similar results for both DPSCs and SHED. The cells showed positive expression for pluripotent markers, ectodermal markers and mesodermal markers. CONCLUSION: The study demonstrated that stem cells existed in human deciduous and permanent pulp tissue. The stem cells present in deciduous permanent pulp tissue can be isolated, cultivated and expanded in vitro. Both DPSCs and SHED show almost a similar expression pattern profile for variety of antigens tested. Further studies should include analysis of diverse cell populations to elucidate their potential to differentiate into various cell types followed by in vivo studies in animals and humans.

scholarly journals VEGFR1 primes a unique cohort of dental pulp stem cells for vasculogenic differentiation

2021 ◽  
Vol 41 ◽  
pp. 332-344
Author(s):  
MT Bergamo ◽  
◽  
Z Zhang ◽  
TM Oliveira ◽  
JE Nör
Keyword(s):  
Stem Cells ◽  
Dental Pulp ◽  
Vascular Endothelial Cells ◽  
Dental Pulp Stem Cells ◽  
Deciduous Teeth ◽  
Microvascular Endothelial Cell ◽  
Vascular Endothelial ◽  
Pulp Tissue ◽  
Immunodeficient Mice

Dental pulp stem cells (DPSCs) constitute a unique group of cells endowed with multipotency, self-renewal, and capacity to regenerate the dental pulp tissue. While much has been learned about these cells in recent years, it is still unclear if each DPSC is multipotent or if unique sub-populations of DPSCs are “primed” to undergo specific differentiation paths. The purpose of the present study was to define whether a sub-population of DPSCs was uniquely primed to undergo vasculogenic differentiation. Permanent-tooth DPSCs or stem cells from human exfoliated deciduous teeth (SHED) were flow-sorted for vascular endothelial growth factor receptor 1 (VEGFR1) and exposed to vasculogenic differentiation medium, i.e., Microvascular-Endothelial-Cell-Growth-Medium-2-BulletKit™ supplemented with 50 ng/mL rhVEGF165 in the presence of 0 or 25 μg/mL anti-human VEGF antibody (bevacizumab; Genentech). In addition, sorted SHED (i.e., VEGFR1high or VEGFR1low) were seeded in biodegradable scaffolds and transplanted into the subcutaneous space of immunodeficient mice. Despite proliferating at a similar rate, VEGFR1high generated more in vitro sprouts than VEGFR1low cells (p < 0.05). Blockade of VEGF signaling with bevacizumab inhibited VEGFR1high-derived sprouts, demonstrating specificity of responses. Similarly, VEGFR1high SHED generated more blood vessels when transplanted into murine hosts than VEGFR1low cells (p < 0.05). Collectively, these data demonstrated that DPSCs contain a unique sub-population of cells defined by high VEGFR1 expression that are primed to differentiate into vascular endothelial cells. These data raise the possibility of purifying stem cells with high vasculogenic potential for regeneration of vascularized tissues or for vascular engineering in the treatment of ischemic conditions.

Stem Cells Derived from Human Exfoliated Deciduous teeth [shed] in Neuronal Disorders: A Review

2020 ◽  
Vol 16 ◽  
Author(s):  
Minu Anoop ◽  
Indrani Datta
Keyword(s):  
Stem Cells ◽  
Neurodegenerative Diseases ◽  
Dental Pulp ◽  
Therapeutic Potential ◽  
Permanent Teeth ◽  
Deciduous Teeth ◽  
Proliferative Potential ◽  
Pulp Tissue ◽  
Human Exfoliated Deciduous Teeth

: Most conventional treatments for neurodegenerative diseases fail due to their focus on neuroprotection rather than neurorestoration. Stem cell‐based therapies are becoming a potential treatment option for neurodegenerative diseases as they can home in, engraft, differentiate and produce factors for CNS recovery. Stem cells derived from human dental pulp tissue differ from other sources of mesenchymal stem cells due to their embryonic neural crest origin and neurotrophic property. These include both dental pulp stem cells [DPSCs] from dental pulp tissues of human permanent teeth and stem cells from human exfoliated deciduous teeth [SHED]. SHED offer many advantages over other types of MSCs such as good proliferative potential, minimal invasive procurement, neuronal differentiation and neurotrophic capacity, and negligible ethical concerns. The therapeutic potential of SHED is attributed to the paracrine action of extracellularly released secreted factors, specifically the secretome, of which exosomes is a key component. SHED and its conditioned media can be effective in neurodegeneration through multiple mechanisms, including cell replacement, paracrine effects, angiogenesis, synaptogenesis, immunomodulation, and apoptosis inhibition, and SHED exosomes offer an ideal refined bed-to-bench formulation in neurodegenerative disorders. However, in spite of these advantages, there are still some limitations of SHED exosome therapy, such as the effectiveness of long-term storage of SHED and their exosomes, the development of a robust GMP-grade manufacturing protocol, optimization of the route of administration, and evaluation of the efficacy and safety in humans. In this review, we have addressed the isolation, collection and properties of SHED along with its therapeutic potential on in vitro and in vivo neuronal disorder models as evident from the published literature.

Isolation and Characterization of Dental Pulp Stem Cells from a Patient with Papillon–Lefèvre Syndrome

2013 ◽  
Vol 39 (1) ◽  
pp. 31-38 ◽  
Author(s):  
Pakize Neslihan Taşlı ◽  
Sıdıka Tapşın ◽  
Sezin Demirel ◽  
Mehmet Emir Yalvaç ◽  
Serap Akyuz ◽  
...  
Keyword(s):  
Stem Cells ◽  
Dental Pulp ◽  
Dental Pulp Stem Cells ◽  
Isolation And Characterization

scholarly journals Hypoxia-inducible factor 1α induces osteo/odontoblast differentiation of human dental pulp stem cells via Wnt/β-catenin transcriptional cofactor BCL9

2022 ◽  
Vol 12 (1) ◽  
Author(s):  
Shion Orikasa ◽  
Nobuyuki Kawashima ◽  
Kento Tazawa ◽  
Kentaro Hashimoto ◽  
Keisuke Sunada-Nara ◽  
...  
Keyword(s):  
Stem Cells ◽  
Dental Pulp ◽  
Target Genes ◽  
Hypoxia Inducible Factor ◽  
Dental Pulp Stem Cells ◽  
Pulp Tissue ◽  
Odontoblast Differentiation ◽  
Hypoxia Inducible Factor 1Α ◽  
Hypoxic Conditions

AbstractAccelerated dental pulp mineralization is a common complication in avulsed/luxated teeth, although the mechanisms underlying this remain unclear. We hypothesized that hypoxia due to vascular severance may induce osteo/odontoblast differentiation of dental pulp stem cells (DPSCs). This study examined the role of B-cell CLL/lymphoma 9 (BCL9), which is downstream of hypoxia-inducible factor 1α (HIF1α) and a Wnt/β-catenin transcriptional cofactor, in the osteo/odontoblastic differentiation of human DPSCs (hDPSCs) under hypoxic conditions. hDPSCs were isolated from extracted healthy wisdom teeth. Hypoxic conditions and HIF1α overexpression induced significant upregulation of mRNAs for osteo/odontoblast markers (RUNX2, ALP, OC), BCL9, and Wnt/β-catenin signaling target genes (AXIN2, TCF1) in hDPSCs. Overexpression and suppression of BCL9 in hDPSCs up- and downregulated, respectively, the mRNAs for AXIN2, TCF1, and the osteo/odontoblast markers. Hypoxic-cultured mouse pulp tissue explants showed the promotion of HIF1α, BCL9, and β-catenin expression and BCL9-β-catenin co-localization. In addition, BCL9 formed a complex with β-catenin in hDPSCs in vitro. This study demonstrated that hypoxia/HIF1α-induced osteo/odontoblast differentiation of hDPSCs was partially dependent on Wnt/β-catenin signaling, where BCL9 acted as a key mediator between HIF1α and Wnt/β-catenin signaling. These findings may reveal part of the mechanisms of dental pulp mineralization after traumatic dental injury.

scholarly journals Isolation and culture of dental pulp stem cells from permanent and deciduous teeth

2019 ◽  
Vol 35 (4) ◽  
Author(s):  
Shagufta Naz ◽  
Farhan Raza Khan ◽  
Raheela Rahmat Zohra ◽  
Sahreena Salim Lakhundi ◽  
Mehwish Sagheer Khan ◽  
...  
Keyword(s):  
Stem Cells ◽  
Dental Pulp ◽  
Dental Pulp Stem Cells ◽  
Permanent Teeth ◽  
Deciduous Teeth ◽  
Tooth Pulp ◽  
Human Teeth ◽  
Creative Commons ◽  
Calcified Tissue

Objective: To isolate dental pulp mesenchymal stem cells (MSCs) from non-infected human permanent and deciduous teeth. Methods: It was an in-vitro experimental study. Human teeth were collected from 13 apparently healthy subjects including nine adults and four children. After decoronation dental pulps were extirpated from teeth and cultured via explant method in a stem cell defined media. Data was analyzed by descriptive statistics. Results: As above MSCs emerged exhibiting fibroblast-like morphology. In vitro culture was positive for 100% (9/9) and 75% (3/4) of the permanent and deciduous teeth respectively. First cell appeared from deciduous teeth pulp in 10±6.2 days while permanent teeth pulp took 12.4±3.7 days. Together, 26.6±3.6 and 24.5±3.5 days were required for permanent and deciduous tooth pulp stem cells to be ready for further assays. Conclusions: The protocol we developed is easy and consistent and can be used to generate reliable source of MScs for engineering of calcified and non-calcified tissue for regenerative medicine approaches. doi: https://doi.org/10.12669/pjms.35.4.540 How to cite this:Naz S, Khan FR, Zohra RR, Lakhundi SS, Khan MS, Mohammed N, et al. Isolation and culture of dental pulp stem cells from permanent and deciduous teeth. Pak J Med Sci. 2019;35(4):---------. doi: https://doi.org/10.12669/pjms.35.4.540 This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/3.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

scholarly journals In vitro histomorphometric comparison of dental pulp tissue in different teeth

PeerJ ◽  
2019 ◽  
Vol 7 ◽  
pp. e8212
Author(s):  
Marytere Guerrero-Jiménez ◽  
Geovanny I. Nic-Can ◽  
Nelly Castro-Linares ◽  
Fernando Javier Aguilar-Ayala ◽  
Michel Canul-Chan ◽  
...  
Keyword(s):  
Stem Cells ◽  
Cell Density ◽  
Dental Pulp ◽  
Pearson Correlation ◽  
Third Molar ◽  
Permanent Teeth ◽  
Deciduous Teeth ◽  
Pulp Tissue ◽  
Valuable Source

Background Dental pulp (DP) represents an accessible and valuable source promising of stem cells for clinical application. However, there are some disadvantages associated with the isolation of dental pulp stem cells (DPSCs), which include the size and weight of the pulp tissue needed to yield sufficient cells for culturing in vitro. Therefore, the objective of this study was to compare in vitro histomorphometry of DP from permanent (premolars, third molar), supernumerary and deciduous teeth of patients between 5 and 25 years old with regards to weight, length, width and the cell density in the four regions of the DP in order to obtain quantitative parameters in a tissue that represents a valuable source of stem cells. Methods DPs were obtained from 10 central incisors deciduous, 20 permanent teeth (10 premolars, 10 third molars) and 10 supernumeraries (six mesiodents and four inferior premolar shapes). The pulps were carefully removed, and the entire tissue was weighed. The pulp length and the width were measured with a digital Vernier caliper. The cellular density analysis was performed according to the four regions of the DP (coronal, cervical, medial and apical) in histological slides using photography and the ImageJ® program for quantification. Results The Pearson correlation test revealed that DP weight among different types of teeth is correlated with age in male patients. A significant positive correlation was noted between length and width of the DP with age in both genders. The mean DP weight for supernumerary and third molar teeth was greater than deciduous and premolar teeth. Finally, the histological analysis showed that the coronal and apical portions of DP in supernumerary and premolar teeth have the highest cell density. Conclusions The DP of supernumerary teeth has quantitatively the best morphometric parameters and cell density comparable with the quality of DP obtained from deciduous teeth.

Isolation and characterization of dental pulp stem cells from a supernumerary tooth

2008 ◽  
Vol 37 (9) ◽  
pp. 571-574 ◽  
Author(s):  
Anderson Hsien-Cheng Huang ◽  
Yuk-Kwan Chen ◽  
Lin-Min Lin ◽  
Tien-Yue Shieh ◽  
Anthony Wing-Sang Chan
Keyword(s):  
Stem Cells ◽  
Dental Pulp ◽  
Dental Pulp Stem Cells ◽  
Supernumerary Tooth ◽  
Isolation And Characterization

scholarly journals Insight into the Role of Dental Pulp Stem Cells in Regenerative Therapy

Biology ◽  
2020 ◽  
Vol 9 (7) ◽  
pp. 160 ◽  
Author(s):  
Shinichiro Yoshida ◽  
Atsushi Tomokiyo ◽  
Daigaku Hasegawa ◽  
Sayuri Hamano ◽  
Hideki Sugii ◽  
...  
Keyword(s):  
Stem Cells ◽  
Tissue Regeneration ◽  
Dental Pulp ◽  
High Growth ◽  
Dental Pulp Stem Cells ◽  
Permanent Teeth ◽  
Deciduous Teeth ◽  
Pulp Tissue ◽  
Differentiation Potential ◽  
Cell Based Therapy

Mesenchymal stem cells (MSCs) have the capacity for self-renewal and multilineage differentiation potential, and are considered a promising cell population for cell-based therapy and tissue regeneration. MSCs are isolated from various organs including dental pulp, which originates from cranial neural crest-derived ectomesenchyme. Recently, dental pulp stem cells (DPSCs) and stem cells from human exfoliated deciduous teeth (SHEDs) have been isolated from dental pulp tissue of adult permanent teeth and deciduous teeth, respectively. Because of their MSC-like characteristics such as high growth capacity, multipotency, expression of MSC-related markers, and immunomodulatory effects, they are suggested to be an important cell source for tissue regeneration. Here, we review the features of these cells, their potential to regenerate damaged tissues, and the recently acquired understanding of their potential for clinical application in regenerative medicine.

scholarly journals Taxifolin Protects Dental Pulp Stem Cells under Hypoxia and Inflammation Conditions

2021 ◽  
Vol 30 ◽  
pp. 096368972110344
Author(s):  
Xiaohui Fu ◽  
Yimiao Feng ◽  
Bingyi Shao ◽  
Yanzhen Zhang
Keyword(s):  
Stem Cells ◽  
Dental Pulp ◽  
Annexin V ◽  
Protective Role ◽  
Dental Pulp Stem Cells ◽  
Pulp Tissue ◽  
Inflammatory Conditions ◽  
Human Dental Pulp ◽  
Unique Source

Background: Dental pulp stem cells (DPSCs) are a unique source for future clinical application in dentistry such as periodontology or endodontics. However, DPSCs are prone to apoptosis under abnormal conditions. Taxifolin is a natural flavonoid and possesses many pharmacological activities including anti-hypoxic and anti-inflammatory. We aimed to elucidate the mechanisms of taxifolin protects DPSC under hypoxia and inflammatory conditions. Methods: DPSCs from human dental pulp tissue was purchased from Lonza (cat. no. PT-5025. Basel, Switzerland)) and identified by DPSC’s biomarkers. DPSC differentiation in vitro following the manufacturers’ instructions. ARS staining and Oil red staining verify the efficiency of differentiation in vitro after 2 weeks. The changes of various genes and proteins were identified by Q-PCR and western-blot, respectively. Cell viability was determined by the CCK-8 method, while apoptosis was determined by Annexin V/PI staining. Results: DPSC differentiation in vitro shows that hypoxia and TNF-α synergistically inhibit the survival and osteogenesis of DPSCs. A final concentration of 10 μM Taxifolin can significantly reduce the apoptosis of DPSCs under inflammation and hypoxia conditions. Taxifolin substantially increases carbonic anhydrase IX (CA9) expression but not HIF1a, and inhibitions of CA9 expression nullify the protective role of taxifolin under hypoxia and inflammatory condition. Conclusion: Taxifolin significantly increased the expression of CA9 when it inhibits DPSC apoptosis and taxifolin synergistically to protect DPSCs against apoptosis with CA9 under hypoxia and inflammatory conditions. Taxifolin can be used as a potential drug for clinical treatment of DPSC-related diseases.

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