Software tools for cell culture-related 3D printed structures

Gulyas, Marton; Csiszer, Miklos; Méhes, Előd; Czirók, András

doi:10.1371/journal.pone.0203203

Cited by 28 publications

(26 citation statements)

References 17 publications

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“…To maintain high cell density cultures for several days, we restricted cell spreading. Our cell culture utilized three, 6 mm diameter mini-wells, fusion-deposition (“3D”) printed from polylactic acid (PLA) into standard 35 mm tissue culture dishes (Greiner) 60 . As cells covered only 10% of the culture area, this configuration permitted to achieve high cell densities ( ) with a high medium/cell ratio.…”

Section: Methodsmentioning

confidence: 99%

Multicellular contractility contributes to the emergence of mesothelioma nodules

Tarnoki-Zach

Stockhammer

Isai

et al. 2020

Sci Rep

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Malignant pleural mesothelioma (MPM) has an overall poor prognosis and unsatisfactory treatment options. MPM nodules, protruding into the pleural cavity may have growth and spreading dynamics distinct that of other solid tumors. We demonstrate that multicellular aggregates can develop spontaneously in the majority of tested MPM cell lines when cultured at high cell density. Surprisingly, the nodule-like aggregates do not arise by excessive local cell proliferation, but by myosin II-driven cell contractility. Prominent actin cables, spanning several cells, are abundant both in cultured aggregates and in MPM surgical specimens. We propose a computational model for in vitro MPM nodule development. Such a self-tensioned Maxwell fluid exhibits a pattern-forming instability that was studied by analytical tools and computer simulations. Altogether, our findings may underline a rational for targeting the actomyosin system in MPM.

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Section: Methodsmentioning

confidence: 99%

Multicellular contractility contributes to the emergence of mesothelioma nodules

Tarnoki-Zach

Stockhammer

Isai

et al. 2020

Sci Rep

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“…The integration of nanomaterials in 3D printing technologies is reviewed in [ 19 ], for the fabrication of novel electronic, biomedical, and bioelectronic devices. Custom fabricated, biocompatible 3D-printed plastic structures are presented in [ 20 ], which can control cell spreading area or medium volume, and exhibit excellent optical properties at 50 µL sample volumes. In [ 15 ], a high-performance 3D-printable conducting polymer ink based on PEDOT:PSS, capable of rapid and flexible fabrication of highly conductive microscale structures and devices both in dry and hydrogel states, is presented.…”

Section: 3d-printed Sensors In Cell Culture and Tissue Engineeringmentioning

confidence: 99%

3D-Printed Sensors and Actuators in Cell Culture and Tissue Engineering: Framework and Research Challenges

Pérez

Serrano

Olmo

2020

Sensors

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Three-dimensional printing technologies have been recently proposed to monitor cell cultures and implement cell bioreactors for different biological applications. In tissue engineering, the control of tissue formation is crucial to form tissue constructs of clinical relevance, and 3D printing technologies can also play an important role for this purpose. In this work, we study 3D-printed sensors that have been recently used in cell culture and tissue engineering applications in biological laboratories, with a special focus on the technique of electrical impedance spectroscopy. Furthermore, we study new 3D-printed actuators used for the stimulation of stem cells cultures, which is of high importance in the process of tissue formation and regenerative medicine. Key challenges and open issues, such as the use of 3D printing techniques in implantable devices for regenerative medicine, are also discussed.

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“…For the analysis of stream formation and collective motility, MEPM cells were seeded at low (30/mm 2 ), medium (100/mm 2 ), and high (300/mm 2 ) densities into 3Dprinted 6mm diameter rings on 35mm tissue culture dishes (Gulyas et al 2018). Cultures were live-imaged every 20 minutes with a 4x phase contrast objective.…”

Section: Spontaneous Collective Motility Assaymentioning

confidence: 99%

SPECC1L-deficient palate mesenchyme cells show speed and directionality defect

Goering

Isai

Hall

et al. 2019

Preprint

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Clefts of the lip and/or palate (CL/P) are common anomalies that occur in 1/800 live births. Pathogenic SPECC1L variants identified in patients with rare atypical clefts and syndromic CL/P suggest the gene plays a primary role in face and palate development. We have generated Specc1l gene-trap (Specc1l cGT ) and truncation (Specc1l DC510 ) alleles that cause embryonic or perinatal lethality, respectively.Specc1l cGT/DC510 compound mutants show delayed and abnormal palatal shelf elevation at E14.5. By E15.5, the mutant shelves do elevate and fuse, however, the palatal rugae form abnormally. Palatogenesis requires extensive mesenchymal remodeling, especially during palatal shelf elevation. We posit that this remodeling involves collective movement of neural crest-derived palatal mesenchyme cells. Live time-lapse microscopy was performed to visualize in vitro wound-repair assays with wildtype and SPECC1L-deficient primary mouse embryonic palatal mesenchyme (MEPM) cells. SPECC1L-deficient MEPM cells consistently showed delayed closure in wound-repair assays. To evaluate which features of cellular movement were responsible, we performed automated particle image velocimetry (PIV) and manual cell tracking. The analyses revealed that both cell speed and directionality are disrupted in SPECC1L-deficient cells compared to controls.To determine if primary MEPM cells can move collectively, we assayed stream formation, which is a hallmark of collective movement. Indeed, MEPM cultures displayed correlated movement of neighboring cells. Importantly, correlation length was reduced in SPECC1Ldeficient cultures, consistent with a role for SPECC1L in collective migration.Furthermore, we demonstrated that activation of the PI3K-AKT pathway with the 740Y-P small molecule can rescue the wound-closure delay in SPECC1L-deficient MEPM cells. 3Cell tracking analyses showed that this rescue was due to both increased speed and improved directionality. Altogether, our data showed a novel role for SPECC1L in guided movement through modulation of PI3K-AKT signaling.

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Software tools for cell culture-related 3D printed structures

Cited by 28 publications

References 17 publications

Multicellular contractility contributes to the emergence of mesothelioma nodules

Multicellular contractility contributes to the emergence of mesothelioma nodules

3D-Printed Sensors and Actuators in Cell Culture and Tissue Engineering: Framework and Research Challenges

SPECC1L-deficient palate mesenchyme cells show speed and directionality defect

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