Fibroblasts maintained in 3 dimensions show a better differentiation state and higher sensitivity to estrogens

Montani, Claudia; Steimberg, Nathalie; Boniotti, Jennifer; Biasiotto, Giorgio; Zanella, Isabella; Diafera, Giuseppe; Biunno, Ida; Caimi, Luigi; Mazzoleni, Giovanna; Lorenzo, Diego Di

doi:10.1016/j.taap.2014.08.021

Cited by 17 publications

(9 citation statements)

References 115 publications

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“…These results are different from those of Barkhordar et al (2002) who showed that activated inflammatory pulpal fibroblasts synthesize much higher amounts of collagen than those from healthy dental pulps. In the same way, Montani et al (2014) indicated that type I collagen was overexpressed in three-dimensional cultures of dermal immortalized murine fibroblasts. However, TNF-a and IL-1b were shown to downregulate the production and mRNA levels of type I collagen in lung fibroblasts (Diaz et al 1993).…”

Section: Discussionmentioning

confidence: 82%

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Expression of factors involved in dental pulp physiopathological processes by nemotic human pulpal fibroblasts

et al. 2017

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Le Clerc J, Tricot-Doleux S, Pellen-Mussi P, P erard M, Jeanne S, P erez F. Expression of factors involved in dental pulp physiopathological processes by nemotic human pulpal fibroblasts. International Endodontic Journal, 51, e94-e106, 2018.Aim To investigate in human dental pulp fibroblasts (HDPF) the expression of factors involved in dental pulp physiopathological processes and in an experimental model of cell activation called nemosis, and to compare the behaviour of pulp cell activation with sound lung fibroblast MRC5, employed as a reference model for nemosis. Methodology Nemotic response was induced in three-dimensional cultures of HDPF and lung fibroblasts. The expressions of molecules involved in physiological (alkaline phosphatase, type I collagen) and in inflammatory processes (IL-6, CXCL8, CCL20, COX-2) were studied using real-time PCR. Concentrations of IL-6 and CXCL8 were analysed during 4 days with ELISA. Nonparametric tests were used to determine statistical differences between groups. Results A significant decrease (P < 0.001) in type I collagen and alkaline phosphatase was observed in MRC5 and HDPF nemotic responses. Although the amounts of mRNA differed between these cell types, there was an increase in CCL20, CXCL8 and COX-2 expression (P < 0.001). Unlike HDPF, MRC5 spheroids displayed significant amounts of IL-6 concentrations and mRNA expression. Notably, increased concentrations of CXCL8 were recorded in all threedimensional cultures compared with monolayers as a function of time (P < 0.05). Conclusion Although the nemotic responses observed were not identical in the pulpal and lung fibroblasts, similarities occurred in the expression of chemokines and cyclooxygenase-2. Nemotic reactions and inflammatory processes in pulp diseases share similarities in terms of the expression of factors. Thus, this in vitro model could constitute a powerful tool to study intercellular relations within the dental pulp and to develop new local treatments to counteract the inflammatory reaction that occurs during pulpitis.

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Section: Discussionmentioning

confidence: 82%

“…In the same way, Montani et al . () indicated that type I collagen was overexpressed in three‐dimensional cultures of dermal immortalized murine fibroblasts. However, TNF‐α and IL‐1β were shown to downregulate the production and mRNA levels of type I collagen in lung fibroblasts (Diaz et al .…”

Section: Discussionmentioning

confidence: 99%

Expression of factors involved in dental pulp physiopathological processes by nemotic human pulpal fibroblasts

et al. 2017

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“…Differentiation in spheroid culture includes not only conservation of morphogenic capacities and histotypic reorganization but also maintenance of functional activities and gene expression patterns. [19][20][21][22][23][24] Traditional methods for tumor spheroid generation include hanging drop, 25 liquid overlaying, spinner flasks, 26 rotary cell culture systems, 27 poly-2-hydroxyethyl methacrylate, low binding plates, 28 gel/ matrix based culture, 29,30 microencapsulation, 31 polymeric scaffolds, 30,32 and micropatterned plates. 8,[33][34][35] However, these methods are frequently associated with poor reproducibility, scalability, adaptability, and lack high-throughput formats amendable to automation and drug screening.…”

Section: Introductionmentioning

confidence: 99%

Generation of Homogenous Three-Dimensional Pancreatic Cancer Cell Spheroids Using an Improved Hanging Drop Technique

Ware

Colbert

Keshishian

et al. 2016

Tissue Engineering Part C: Methods

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In vitro characterization of tumor cell biology or of potential anticancer drugs is usually performed using tumor cell lines cultured as a monolayer. However, it has been previously shown that three-dimensional (3D) organization of the tumor cells is important to provide insights on tumor biology and transport of therapeutics. Several methods to create 3D tumors in vitro have been proposed, with hanging drop technique being the most simple and, thus, most frequently used. However, in many cell lines this method has failed to form the desired 3D tumor structures. The aim of this study was to design and test an easy-to-use and highly reproducible modification of the hanging drop method for tumor sphere formation by adding methylcellulose polymer. Most pancreatic cancer cells do not form cohesive and manageable spheres when the original hanging drop method is used, thus we investigated these cell lines for our modified hanging drop method. The spheroids produced by this improved technique were analyzed by histology, light microscopy, immunohistochemistry, and scanning electron microscopy. Results show that using the proposed simple method; we were able to produce uniform spheroids for all five of the tested human pancreatic cancer cell lines; Panc-1, BxPC-3, Capan-1, MiaPaCa-2, and AsPC-1. We believe that this method can be used as a reliable and reproducible technique to make 3D cancer spheroids for use in tumor biology research and evaluation of therapeutic responses, and for the development of bio-artificial tissues.

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“…Cell culture was established from a suspension of 1 × 10 6 cells/ml in RPMI 1640 medium, supplemented with 10%FBS, L‐glutamine and penicillin/streptomycin and maintained at 37 °C, in a humidified atmosphere with 5% CO 2 . The rotation speed of the culture vessel (7.5 rpm) is known to furnish an optimal hydrodynamic microenvironment . Twenty‐four hours after seeding, the culture medium was replaced by the chemicals to be tested, dissolved in complete culture medium.…”

Section: Methodsmentioning

confidence: 99%

Paclitaxel‐releasing mesenchymal stromal cells inhibit the growth of multiple myeloma cells in a dynamic 3D culture system

Bonomi

Steimberg

Benetti

et al. 2016

Hematological Oncology

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Multiple myeloma is an aggressive tumour able to suppress osteoblastogenesis probably mediated by bone marrow mesenchymal stromal cells (BM-MSCs) that can also support plasma cell growth/survival. The use of MSCs for multiple myeloma therapy is a controversial topic because of the contradictory results on the capacity of MSCs to inhibit or to promote cancer growth. Our previous studies demonstrated that MSCs could be loaded with Paclitaxel (PTX) and used to deliver the drug in situ in amount affecting tumour growth (in vitro and in vivo). Therefore, independently on the discussed action of MSCs in myeloma, MSCs could represent a 'trojan horse' to vehicle and deliver anti-tumour agents into bone marrow. This study confirms, by an in vitro 3D dynamic culture system, that PTX loaded BM-MSCs (PTXr-MSCs) are active on the proliferation of RPMI 8226, a human myeloma cell line. Our results demonstrated a dramatic suppression of myeloma cell growth by PTXr-MSCs, suggesting that drug loaded MSCs could be a tool to deliver drug into the bone marrow. Drug releasing MSCs provide a therapeutic approach to potentiate the existing treatments against a very aggressive malignancy as multiple myeloma. Copyright © 2016 John Wiley & Sons, Ltd.

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Fibroblasts maintained in 3 dimensions show a better differentiation state and higher sensitivity to estrogens

Cited by 17 publications

References 115 publications

Expression of factors involved in dental pulp physiopathological processes by nemotic human pulpal fibroblasts

Expression of factors involved in dental pulp physiopathological processes by nemotic human pulpal fibroblasts

Generation of Homogenous Three-Dimensional Pancreatic Cancer Cell Spheroids Using an Improved Hanging Drop Technique

Paclitaxel‐releasing mesenchymal stromal cells inhibit the growth of multiple myeloma cells in a dynamic 3D culture system

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