Manufacturing of dental pulp cell-based products from human third molars: current strategies and future investigations

Ducret, Maxime; Fabre, Hugo; Degoul, Olivier; Atzeni, Gianluigi; McGuckin, Colin; Forraz, Nico; Alliot‐Licht, Brigitte; Malléin-Gerin, Frédéric; Perrier‐Groult, Emeline; Farges, Jean‐Christophe

doi:10.3389/fphys.2015.00213

Cited by 36 publications

(40 citation statements)

References 83 publications

(123 reference statements)

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“…A recent report [185] described manufacturing strategies of DPSC-based ATMPs to improve safety, efficacy, and consistency of their GMP production. The authors proposed the use of impacted third molars of young healthy donors between 5 and 7 Nolla's developmental stage (i.e., from complete crown upto one third of root completed) as ideal dental MSC sources.…”

Section: Establishment Of Clinical-grade Dental Mscs and Challengementioning

confidence: 99%

Stem Cells of Dental Origin: Current Research Trends and Key Milestones towards Clinical Application

Bakopoulou

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2016

Stem Cells International

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Dental Mesenchymal Stem Cells (MSCs), including Dental Pulp Stem Cells (DPSCs), Stem Cells from Human Exfoliated Deciduous teeth (SHED), and Stem Cells From Apical Papilla (SCAP), have been extensively studied using highly sophisticated in vitro and in vivo systems, yielding substantially improved understanding of their intriguing biological properties. Their capacity to reconstitute various dental and nondental tissues and the inherent angiogenic, neurogenic, and immunomodulatory properties of their secretome have been a subject of meticulous and costly research by various groups over the past decade. Key milestone achievements have exemplified their clinical utility in Regenerative Dentistry, as surrogate therapeutic modules for conventional biomaterial-based approaches, offering regeneration of damaged oral tissues instead of simply “filling the gaps.” Thus, the essential next step to validate these immense advances is the implementation of well-designed clinical trials paving the way for exploiting these fascinating research achievements for patient well-being: the ultimate aim of this ground breaking technology. This review paper presents a concise overview of the major biological properties of the human dental MSCs, critical for the translational pathway “from bench to clinic.”

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Section: Establishment Of Clinical-grade Dental Mscs and Challengementioning

confidence: 99%

Stem Cells of Dental Origin: Current Research Trends and Key Milestones towards Clinical Application

Bakopoulou

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2016

Stem Cells International

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“…Dental and craniofacial research currently explores a variety of cell-based and tissue engineering protocols to be used as alternatives to classical therapies that aimed at repairing/regenerating damaged tissues (Ducret et al, 2015a; Gorin et al, 2016). In particular, studies have demonstrated that mesenchymal stromal/stem cells (MSCs) are suitable to these protocols because of their high expansion ability and differentiation potential both in culture and animal models.…”

Section: Introductionmentioning

confidence: 99%

“…We recently isolated and easily amplified in culture a population of MSCs from the DP of human developing third molars with a medicinal manufacturing approach (Ducret et al, 2015b). We showed by using flow cytometry that all cells of this population expressed the mesenchymal cell markers CD10, CD13, CD29, CD44, CD90, CD105, and CD166 in vitro , but did not express the hematopoietic markers CD14, CD34, CD45, CD79a, and HLA-DR or the endothelial cell/leucocyte marker CD31 (Ducret et al, 2015a,b; Ducret et al, 2016). Stro-1 and CD271 were expressed by a very low number (≤ 1%) of cultured DP-MSCs, whereas CD146 and MSCA-1 were expressed by about 40 and 15% of DP-MSCs, respectively.…”

Section: Introductionmentioning

confidence: 99%

Immunophenotyping Reveals the Diversity of Human Dental Pulp Mesenchymal Stromal Cells In vivo and Their Evolution upon In vitro Amplification

Ducret

Fabre

Degoul³

et al. 2016

Front. Physiol.

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Mesenchymal stromal/stem cells (MSCs) from human dental pulp (DP) can be expanded in vitro for cell-based and regenerative dentistry therapeutic purposes. However, their heterogeneity may be a hurdle to the achievement of reproducible and predictable therapeutic outcomes. To get a better knowledge about this heterogeneity, we designed a flow cytometric strategy to analyze the phenotype of DP cells in vivo and upon in vitro expansion with stem cell markers. We focused on the CD31− cell population to exclude endothelial and leukocytic cells. Results showed that the in vivo CD31− DP cell population contained 1.4% of CD56+, 1.5% of CD146+, 2.4% of CD271+ and 6.3% of MSCA-1+ cells but very few Stro-1+ cells (≤ 1%). CD56+, CD146+, CD271+, and MSCA-1+ cell subpopulations expressed various levels of these markers. CD146+MSCA-1+, CD271+MSCA-1+, and CD146+CD271+ cells were the most abundant DP-MSC populations. Analysis of DP-MSCs expanded in vitro with a medicinal manufacturing approach showed that CD146 was expressed by about 50% of CD56+, CD271+, MSCA-1+, and Stro-1+ cells, and MSCA-1 by 15–30% of CD56+, CD146+, CD271+, and Stro-1+ cells. These ratios remained stable with passages. CD271 and Stro-1 were expressed by <1% of the expanded cell populations. Interestingly, the percentage of CD56+ cells strongly increased from P1 (25%) to P4 (80%) both in all sub-populations studied. CD146+CD56+, MSCA-1+CD56+, and CD146+MSCA-1+ cells were the most abundant DP-MSCs at the end of P4. These results established that DP-MSCs constitute a heterogeneous mixture of cells in pulp tissue in vivo and in culture, and that their phenotype is modified upon in vitro expansion. Further studies are needed to determine whether co-expression of specific MSC markers confers DP cells specific properties that could be used for the regeneration of human tissues, including the dental pulp, with standardized cell-based medicinal products.

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“…Explant based isolation could also be easily adapted to xeno‐free isolation and expansion of DPSCs on a clinical scale, which would further simplify regulatory approvals (Ducret et al, ). Further expansion of cells derived by sustained explant technique showed identical surface marker profiles in all cultures, validating the MSC‐phenotype.…”

Section: Resultsmentioning

confidence: 99%

Long term explant culture for harvesting homogeneous population of human dental pulp stem cells

Patil

Kharat

Kulkarni

et al. 2018

Cell Biology International

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Dental pulp stem cells have emerged as a preferred source of mesenchymal stem cells, because of its easy availability and high stem cell content. Dental pulp is a specific fibrous tissue that contains heterogeneous populations of odontoblasts, fibroblasts, pericytes, progenitors, stem cells, leukocytes and neuronal cells. In this study, we propose sustained explant culture as a simple, economical and efficient process to isolate dental pulp stem cells from human Dental pulp Tissue. Historically explant cultures were used to get fibroblast cells from embryonic chick heart using plasma clot cultures. The subculture was performed by lifting mother explant (original explant) and grafting it in a new plasma clot. We modified this age old technique to suit the modern times. Here we demonstrate for the first time that the mother explant (E0) of human dental pulp tissue could be sub‐cultured consecutively seven times (E7) without displacement. This technique is highly reproducible and permits growth and proliferation of dental pulp stem cells yielding an enriched homogeneous mesenchymal stem cells population in the first passage itself as revealed by surface marker expression. These dental pulp stem cells exhibit differentiation into adipogenic, chondrogenic and osteogenic lineage revealing their mesenchymal stem cell nature. We propose that dental pulp stem cells isolated by sustained explant culture are phenotypically and functionally comparable to those obtained by enzymatic method. It is a simple, inexpensive and gentle method, which may be preferred over the conventional techniques for obtaining stem cells from other tissue sources as well especially in cases of limited starting material.

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Manufacturing of dental pulp cell-based products from human third molars: current strategies and future investigations

Cited by 36 publications

References 83 publications

Stem Cells of Dental Origin: Current Research Trends and Key Milestones towards Clinical Application

Stem Cells of Dental Origin: Current Research Trends and Key Milestones towards Clinical Application

Immunophenotyping Reveals the Diversity of Human Dental Pulp Mesenchymal Stromal Cells In vivo and Their Evolution upon In vitro Amplification

Long term explant culture for harvesting homogeneous population of human dental pulp stem cells

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