scholarly journals Culturing and Scaling up Stem Cells of Dental Pulp Origin Using Microcarriers

Polymers ◽  
2021 ◽  
Vol 13 (22) ◽  
pp. 3951
Author(s):  
Anna Földes ◽  
Hajnalka Reider ◽  
Anita Varga ◽  
Krisztina S. Nagy ◽  
Katalin Perczel-Kovach ◽  
...  
Keyword(s):  
Stem Cells ◽  
Dental Pulp ◽  
Glucose Consumption ◽  
Culture Conditions ◽  
Two Photon ◽  
Dynamic Culture ◽  
Shake Flasks ◽  
Electrode Method ◽  
Neural Crest Origin ◽  
Alamarblue Assay

Ectomesenchymal stem cells derived from the dental pulp are of neural crest origin, and as such are promising sources for cell therapy and tissue engineering. For safe upscaling of these cells, microcarrier-based culturing under dynamic conditions is a promising technology. We tested the suitability of two microcarriers, non-porous Cytodex 1 and porous Cytopore 2, for culturing well characterized dental pulp stem cells (DPSCs) using a shake flask system. Human DPSCs were cultured on these microcarriers in 96-well plates, and further expanded in shake flasks for upscaling experiments. Cell viability was measured using the alamarBlue assay, while cell morphology was observed by conventional and two-photon microscopies. Glucose consumption of cells was detected by the glucose oxidase/Clark-electrode method. DPSCs adhered to and grew well on both microcarrier surfaces and were also found in the pores of the Cytopore 2. Cells grown in tissue culture plates (static, non-shaking conditions) yielded 7 × 105 cells/well. In shake flasks, static preincubation promoted cell adhesion to the microcarriers. Under dynamic culture conditions (shaking) 3 × 107 cells were obtained in shake flasks. The DPSCs exhausted their glucose supply from the medium by day seven even with partial batch-feeding. In conclusion, both non-porous and porous microcarriers are suitable for upscaling ectomesenchymal DPSCs under dynamic culture conditions.

scholarly journals Engineering large cartilage tissues using dynamic bioreactor culture at defined oxygen conditions

2018 ◽  
Vol 9 ◽  
pp. 204173141775371 ◽  
Author(s):  
Andrew C Daly ◽  
Binulal N Sathy ◽  
Daniel J Kelly
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Stem Cell ◽  
Mesenchymal Stem Cell ◽  
Culture Conditions ◽  
Cartilage Tissue ◽  
Bioreactor Culture ◽  
Dynamic Culture ◽  
Oxygen Conditions ◽  
Static Conditions

Mesenchymal stem cells maintained in appropriate culture conditions are capable of producing robust cartilage tissue. However, gradients in nutrient availability that arise during three-dimensional culture can result in the development of spatially inhomogeneous cartilage tissues with core regions devoid of matrix. Previous attempts at developing dynamic culture systems to overcome these limitations have reported suppression of mesenchymal stem cell chondrogenesis compared to static conditions. We hypothesize that by modulating oxygen availability during bioreactor culture, it is possible to engineer cartilage tissues of scale. The objective of this study was to determine whether dynamic bioreactor culture, at defined oxygen conditions, could facilitate the development of large, spatially homogeneous cartilage tissues using mesenchymal stem cell laden hydrogels. A dynamic culture regime was directly compared to static conditions for its capacity to support chondrogenesis of mesenchymal stem cells in both small and large alginate hydrogels. The influence of external oxygen tension on the response to the dynamic culture conditions was explored by performing the experiment at 20% O2 and 3% O2. At 20% O2, dynamic culture significantly suppressed chondrogenesis in engineered tissues of all sizes. In contrast, at 3% O2 dynamic culture significantly enhanced the distribution and amount of cartilage matrix components (sulphated glycosaminoglycan and collagen II) in larger constructs compared to static conditions. Taken together, these results demonstrate that dynamic culture regimes that provide adequate nutrient availability and a low oxygen environment can be employed to engineer large homogeneous cartilage tissues. Such culture systems could facilitate the scaling up of cartilage tissue engineering strategies towards clinically relevant dimensions.

Photobiomodulation of inflamed dental pulp stem cells under different nutritional conditions

2021 ◽  
Author(s):  
Seyedeh Sareh Hendi ◽  
Leila Gholami ◽  
Massoud Saidijam ◽  
Roghayeh Mahmoudi ◽  
Ali Asghar Arkian ◽  
...  
Keyword(s):  
Gene Expression ◽  
Stem Cells ◽  
Cell Proliferation ◽  
Laser Irradiation ◽  
Dental Pulp ◽  
Culture Conditions ◽  
Dental Pulp Stem Cells ◽  
Reverse Transcription Pcr ◽  
Nutritional Conditions ◽  
Proliferation And Differentiation

Aim: The present study aimed to investigate photobiomodulation's (PBM) effect on inflamed dental pulp stem cells (IDPSCs) under different nutritional conditions. Methods: Cell proliferation and odontogenic differentiation were evaluated using the MTT assay and real-time quantitative reverse transcription PCR, respectively after laser PBM of cells in 5 or 10% fetal bovine serum (FBS) culture conditions. Results: A significant positive effect of laser irradiation on cell proliferation under both nutritional conditions after 24 and 48 h was observed. DMP-1 gene expression increased in the groups with laser irradiation and 5% FBS. Comparison of gene expression levels in the four groups revealed no statistically significant stimulatory effect. The highest gene expression was observed in the non-laser group with 5% FBS. Conclusion: Further studies are required to obtain an irradiation setup to ideally improve inflamed dental pulp stem cells' proliferation and differentiation.

Phenotypic and functional comparison of optimum culture conditions for upscaling of dental pulp stem cells

2013 ◽  
Vol 37 (2) ◽  
pp. 126-136 ◽  
Author(s):  
Mohammad Mahboob Kanafi ◽  
Rajarshi Pal ◽  
Pawan Kumar Gupta
Keyword(s):  
Stem Cells ◽  
Dental Pulp ◽  
Culture Conditions ◽  
Dental Pulp Stem Cells

scholarly journals Chitlac-coated Thermosets Enhance Osteogenesis and Angiogenesis in a Co-culture of Dental Pulp Stem Cells and Endothelial Cells

Nanomaterials ◽  
2019 ◽  
Vol 9 (7) ◽  
pp. 928 ◽  
Author(s):  
Rapino ◽  
Di Valerio ◽  
Zara ◽  
Gallorini ◽  
Marconi ◽  
...  
Keyword(s):  
Stem Cells ◽  
Endothelial Cells ◽  
Dental Pulp ◽  
Cellular Response ◽  
Cell Types ◽  
Culture Conditions ◽  
Dental Pulp Stem Cells ◽  
Alizarin Red ◽  
Dental Restorations ◽  
Diphenyl Tetrazolium Bromide

Dental pulp stem cells (DPSCs) represent a population of stem cells which could be useful in oral and maxillofacial reconstruction. They are part of the periendothelial niche, where their crosstalk with endothelial cells is crucial in the cellular response to biomaterials used for dental restorations. DPSCs and the endothelial cell line EA.hy926 were co-cultured in the presence of Chitlac-coated thermosets in culture conditions inducing, in turn, osteogenic or angiogenic differentiation. Cell proliferation was evaluated by 3–[4,5–dimethyl–thiazol–2–yl–]–2,5–diphenyl tetrazolium bromide (MTT) assay. DPSC differentiation was assessed by measuring Alkaline Phosphtase (ALP) activity and Alizarin Red S staining, while the formation of new vessels was monitored by optical microscopy. The IL-6 and PGE2 production was evaluated as well. When cultured together, the proliferation is increased, as is the DPSC osteogenic differentiation and EA.hy926 vessel formation. The presence of thermosets appears either not to disturb the system balance or even to improve the osteogenic and angiogenic differentiation. Chitlac-coated thermosets confirm their biocompatibility in the present co-culture model, being capable of improving the differentiation of both cell types. Furthermore, the assessed co-culture appears to be a useful tool to investigate cell response toward newly synthesized or commercially available biomaterials, as well as to evaluate their engraftment potential in restorative dentistry.

Analysis of growth kinetics and colony-forming unit efficiency of human dental pulp stem cells cultured in different media and supplements

2021 ◽  
Vol 16 (7) ◽  
pp. 203-210
Author(s):  
Kumar Chethan ◽  
Shishir Shetty ◽  
Basan Gowda Kurkalli ◽  
Veena Shetty ◽  
Kumar Basavarajappa Mohana
Keyword(s):  
Stem Cells ◽  
Growth Kinetics ◽  
Dental Pulp ◽  
Culture Conditions ◽  
Dental Pulp Stem Cells ◽  
Population Doubling ◽  
Animal Origin ◽  
Dental Tissues ◽  
Colony Forming Unit ◽  
Human Dental Pulp

Dental tissues are considered as ideal autologous sources of multipotent stem cells. Presently, human dental pulp stem cells (DPSCs) are largely being isolated and expanded in media containing fetal bovine serum (FBS). However, the use of FBS has limitations due to its animal origin. Therefore, the present study evaluated the morphology, proliferation rate, population doubling time (PDT) and colony-forming unit fibroblast (CFU-F) efficiency of DPSCs cultured in animal serum-containing medium (SCM) and serumfree medium (SFM) in addition to serum-free culture conditions by supplementing human blood-derivatives such as platelet lysate (PL), fresh frozen plasma (FFP) and umbilical cord blood serum (UCS) at 2.5%, 5% and 7.5% concentrations. Established DPSCs had spindle-shape during primary culture but acquired characteristic fibroblast-like features when cultured in PL, FFP and UCS. DPSCs in SCM, SFM and PL had significantly (P<0.05) higher proliferative potential than those in UCS and FFP and these observations were supported by PDT values. The CFU efficiency of DPSCs was confirmed in all culture conditions with a slightly varied clonogenic potential in blood-derived components. Based on the growth kinetics and CFU ability, it is concluded that PL could be considered as a suitable alternative to FBS for the ex vivo expansion of DPSCs.

scholarly journals CD146 controls the quality of clinical grade mesenchymal stem cells from human dental pulp

2021 ◽  
Vol 12 (1) ◽  
Author(s):  
Lan Ma ◽  
Zhiqing Huang ◽  
Di Wu ◽  
Xiaoxing Kou ◽  
Xueli Mao ◽  
...  
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Dental Pulp ◽  
Culture Conditions ◽  
Proliferation Rate ◽  
Permanent Teeth ◽  
Deciduous Teeth ◽  
Clinical Grade ◽  
Surface Molecule ◽  
Differentiation Potential

Abstract Background Human mesenchymal stem cells from dental pulp (hMSC-DP), including dental pulp stem cells from permanent teeth and exfoliated deciduous teeth, possess unique MSC characteristics such as expression of specific surface molecules and a high proliferation rate. Since hMSC-DP have been applied in numerous clinical studies, it is necessary to establish criteria to evaluate their potency for cell-based therapies. Methods We compared stem cell properties of hMSC-DP at passages 5, 10 and 20 under serum (SE) and serum-free (SF) culture conditions. Cell morphology, proliferation capacity, chromosomal stability, surface phenotypic profiles, differentiation and immunoregulation ability were evaluated. In addition, we assessed surface molecule that regulates hMSC-DP proliferation and immunomodulation. Results hMSC-DP exhibited a decrease in proliferation rate and differentiation potential, as well as a reduced expression of CD146 when cultured under continuous passage conditions. SF culture conditions failed to alter surface marker expression, chromosome stability or proliferation rate when compared to SE culture. SF-cultured hMSC-DP were able to differentiate into osteogenic, adipogenic and neural cells, and displayed the capacity to regulate immune responses. Notably, the expression level of CD146 showed a positive correlation with proliferation, differentiation, and immunomodulation, suggesting that CD146 can serve as a surface molecule to evaluate the potency of hMSC-DP. Mechanistically, we found that CD146 regulates proliferation and immunomodulation of hMSC-DP through the ERK/p-ERK pathway. Conclusion This study indicates that SF-cultured hMSC-DP are appropriate for producing clinical-grade cells. CD146 is a functional surface molecule to assess the potency of hMSC-DP.

scholarly journals Dental Pulp Stem Cells Derived From Adult Human Third Molar Tooth: A Brief Review

2021 ◽  
Vol 9 ◽  
Author(s):  
Ashraf Al Madhoun ◽  
Sardar Sindhu ◽  
Dania Haddad ◽  
Maher Atari ◽  
Rasheed Ahmad ◽  
...  
Keyword(s):  
Stem Cells ◽  
Dental Pulp ◽  
Adult Stem Cells ◽  
Third Molar ◽  
Culture Conditions ◽  
Dental Pulp Stem Cells ◽  
Self Renewal ◽  
Lineage Differentiation ◽  
Physiological Properties ◽  
Underlying Causes

The fields of regenerative medicine and stem cell-based tissue engineering have the potential of treating numerous tissue and organ defects. The use of adult stem cells is of particular interest when it comes to dynamic applications in translational medicine. Recently, dental pulp stem cells (DPSCs) have been traced in third molars of adult humans. DPSCs have been isolated and characterized by several groups. DPSCs have promising characteristics including self-renewal capacity, rapid proliferation, colony formation, multi-lineage differentiation, and pluripotent gene expression profile. Nevertheless, genotypic, and phenotypic heterogeneities have been reported for DPSCs subpopulations which may influence their therapeutic potentials. The underlying causes of DPSCs’ heterogeneity remain poorly understood; however, their heterogeneity emerges as a consequence of an interplay between intrinsic and extrinsic cellular factors. The main objective of the manuscript is to review the current literature related to the human DPSCs derived from the third molar, with a focus on their physiological properties, isolation procedures, culture conditions, self-renewal, proliferation, lineage differentiation capacities and their prospective advances use in pre-clinical and clinical applications.

scholarly journals Optimal culture conditions for neurosphere formation and neuronal differentiation from human dental pulp stem cells

2021 ◽  
Vol 29 ◽  
Author(s):  
Thanasup GONMANEE ◽  
Tawepong ARAYAPISIT ◽  
Kutkao VONGSAVAN ◽  
Chareerut PHRUKSANIYOM ◽  
Hathaitip SRITANAUDOMCHAI
Keyword(s):  
Stem Cells ◽  
Neuronal Differentiation ◽  
Dental Pulp ◽  
Culture Conditions ◽  
Dental Pulp Stem Cells ◽  
Human Dental Pulp
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