Rapid 3D Extrusion of Synthetic Tumor Microenvironments.
Ontology highlight
ABSTRACT:
SUBMITTER: Grolman JM
PROVIDER: S-EPMC4745120 | biostudies-literature | 2015 Oct
REPOSITORIES: biostudies-literature
Publications
Rapid 3D Extrusion of Synthetic Tumor Microenvironments.
Grolman Joshua M JM Zhang Douglas D Smith Andrew M AM Moore Jeffrey S JS Kilian Kristopher A KA
Advanced materials (Deerfield Beach, Fla.) 20150818 37
The spatial positioning of 3D tumor-mimetic microenvironments, containing multiple cell types, is controlled with a simple concentric flow device in a single step. A range of geometric architectures are demonstrated, and the migration of segregated tumor cells and macrophages is explored using drugs that inhibit heterotypic interactions. ...[more]
Similar Datasets
Project description:Three-dimensional organoid constructs serve as increasingly widespread in vitro models for development and disease modeling. Current approaches to recreate morphogenetic processes in vitro rely on poorly controllable and ill-defined matrices, thereby largely overlooking the contribution of biochemical and biophysical extracellular matrix (ECM) factors in promoting multicellular growth and reorganization. Here, we show how defined synthetic matrices can be used to explore the role of the ECM in the development of complex 3D neuroepithelial cysts that recapitulate key steps in early neurogenesis. We demonstrate how key ECM parameters are involved in specifying cytoskeleton-mediated symmetry-breaking events that ultimately lead to neural tube-like patterning along the dorsal-ventral (DV) axis. Such synthetic materials serve as valuable tools for studying the discrete action of extrinsic factors in organogenesis, and allow for the discovery of relationships between cytoskeletal mechanobiology and morphogenesis.
| S-EPMC5098636 | biostudies-literature
Project description:Bioinks with shear-thinning/rapid solidification properties and strong mechanics are usually needed for the bioprinting of three-dimensional (3D) cell-laden constructs. As such, it remains challenging to generate soft constructs from bioinks at low concentrations that are favorable for cellular activities. Herein, we report a strategy to fabricate cell-laden constructs with tunable 3D microenvironments achieved by bioprinting of gelatin methacryloyl (GelMA)/alginate core/sheath microfibers, where the alginate sheath serves as a template to support and confine the GelMA pre-hydrogel in the core during the extrusion process, allowing for subsequent UV crosslinking. This novel strategy minimizes the bioprinting requirements for the core bioink, and facilitates the fabrication of cell-laden GelMA constructs at low concentrations. We first showed the capability of generating various alginate hollow microfibrous constructs using a coaxial nozzle setup, and verified the diffusibility and perfusability of the bioprinted hollow structures that are important for the tissue engineering applications. More importantly, the hollow alginate microfibers were then used as templates for generating cell-laden GelMA constructs with soft microenvironments, by using GelMA pre-hydrogel as the bioink for the core phase during bioprinting. As such, GelMA constructs at extremely low concentrations (<2.0%) could be extruded to effectively support cellular activities including proliferation and spreading for various cell types. We believe that our strategy is likely to provide broad opportunities in bioprinting of 3D constructs with cell-favorable microenvironments for applications in tissue engineering and pharmaceutical screening.
| S-EPMC5837947 | biostudies-literature
Project description:The topographic features of the extracelluar matrix (ECM) lay the foundation for cellular behavior. A novel biofabrication method using a digital-mirror device (DMD), called dynamic optical projection stereolithography (DOPsL) is demonstrated. This robust and versatile platform can generate complex biomimetic scaffolds within seconds. Such 3D scaffolds have promising potentials for studying cell interactions with microenvironments in vitro and in vivo.
| S-EPMC3789064 | biostudies-literature
Project description:Background: Development of new medicines is a lengthy process with high risk of failure since drug efficacy measured in vitro is difficult to confirm in vivo. Intended to add a new tool aiding drug discovery, the MOAB-NICHOID device was developed: a miniaturized optically accessible bioreactor (MOAB) housing the 3D engineered scaffold NICHOID. The aim of our study was to characterize the microflow through the 3D nichoid microenvironment hosted in the MOAB-NICHOID device. Methods: We used computational fluid dynamics (CFD) simulations to compute the flow field inside a very fine grid resembling the scaffold microenvironment. Results: The microflow inside the multi-array of nichoid blocks is fed and locally influenced by the mainstream flow developed in the perfusion chamber of the device. Here we have revealed a low velocity, complex flow field with secondary, backward, or local recirculation micro-flows induced by the intricate architecture of the nichoid scaffold. Conclusion: Knowledge of the microenvironment inside the 3D nichoids allows planning of cell experiments, to regulate the transport of cells towards the scaffold substrate during seeding or the spatial delivery of nutrients and oxygen which affects cell growth and viability.
| S-EPMC8718690 | biostudies-literature
Project description:The development of 3D in vitro models capable of recapitulating native tumor microenvironments could improve the translatability of potential anticancer drugs and treatments. Here, 3D bioprinting techniques are used to build tumor constructs via precise placement of living cells, functional biomaterials, and programmable release capsules. This enables the spatiotemporal control of signaling molecular gradients, thereby dynamically modulating cellular behaviors at a local level. Vascularized tumor models are created to mimic key steps of cancer dissemination (invasion, intravasation, and angiogenesis), based on guided migration of tumor cells and endothelial cells in the context of stromal cells and growth factors. The utility of the metastatic models for drug screening is demonstrated by evaluating the anticancer efficacy of immunotoxins. These 3D vascularized tumor tissues provide a proof-of-concept platform to i) fundamentally explore the molecular mechanisms of tumor progression and metastasis, and ii) preclinically identify therapeutic agents and screen anticancer drugs.
| S-EPMC6996245 | biostudies-literature
Project description:The cancer microenvironment, which incorporates interactions with stromal cells, extracellular matrix (ECM), and other tumor cells in a 3-dimensional (3D) context, has been implicated in every stage of cancer development, including growth of the primary tumor, metastatic spread, and response to treatment. Our understanding of the tumor microenvironment and our ability to develop new therapies would greatly benefit from tools that allow us to systematically probe microenvironmental cues within a 3D context. Here, we leveraged recent advances in microfluidic technology to develop a platform for high-throughput fabrication of tunable cellular microniches ("microtissues") that allow us to probe tumor cell response to a range of microenvironmental cues, including ECM, soluble factors, and stromal cells, all in 3D. We further combine this tunable microniche platform with rapid, flow-based population level analysis (n > 500), which permits analysis and sorting of microtissue populations both pre- and post-culture by a range of parameters, including proliferation and homotypic or heterotypic cell density. We used this platform to demonstrate differential responses of lung adenocarcinoma cells to a selection of ECM molecules and soluble factors. The cells exhibited enhanced or reduced proliferation when encapsulated in fibronectin- or collagen-1-containing microtissues, respectively, and they showed reduced proliferation in the presence of TGF-β, an effect that we did not observe in monolayer culture. We also measured tumor cell response to a panel of drug targets and found, in contrast to monolayer culture, specific sensitivity of tumor cells to TGFβR2 inhibitors, implying that TGF-β has an anti-proliferative affect that is unique to the 3D context and that this effect is mediated by TGFβR2. These findings highlight the importance of the microenvironmental context in therapeutic development and that the platform we present here allows the high-throughput study of tumor response to drugs as well as basic tumor biology in well-defined microenvironmental niches.
| S-EPMC3812960 | biostudies-literature
Project description:Fibroblasts are a heterogeneous group of cells comprising subpopulations that have been found to be activated in the stromal microenvironment that regulates tumor initiation and growth. The underlying mechanisms of such selective activation of fibroblasts are not understood. We propose that the intrinsic geometric heterogeneity of fibroblasts modulates the nuclear mechanotransduction of signals from the microenvironment, resulting in their selective activation. To test this, we developed an engineered 3D fibroblast tumor coculture system and used high resolution images to quantify multiple cell geometry sensitive nuclear morphological and chromatin organizational features. These features were then mapped to activation levels as measured by the nuclear abundance of transcription cofactor, megakaryoblastic leukemia, and protein levels of its target, αSMA. Importantly, our results indicate the presence of activation-"primed" cell geometries that present higher activation levels, which are further enhanced in the presence of stimuli from cancer cells. Further, we show that by enriching the population of activation-primed cell geometric states by either increasing matrix rigidity or micropatterning primed cell shapes, fibroblast activation levels can be increased. Collectively, our results reveal important cellular geometric states that select for fibroblast activation within the heterogenous tumor microenvironment.
| S-EPMC7185960 | biostudies-literature
Project description:Defining the principles of T cell migration in structurally and mechanically complex tumor microenvironments is critical to understanding escape from antitumor immunity and optimizing T cell-related therapeutic strategies. Here, we engineered nanotextured elastic platforms to study and enhance T cell migration through complex microenvironments and define how the balance between contractility localization-dependent T cell phenotypes influences migration in response to tumor-mimetic structural and mechanical cues. Using these platforms, we characterize a mechanical optimum for migration that can be perturbed by manipulating an axis between microtubule stability and force generation. In 3D environments and live tumors, we demonstrate that microtubule instability, leading to increased Rho pathway-dependent cortical contractility, promotes migration whereas clinically used microtubule-stabilizing chemotherapies profoundly decrease effective migration. We show that rational manipulation of the microtubule-contractility axis, either pharmacologically or through genome engineering, results in engineered T cells that more effectively move through and interrogate 3D matrix and tumor volumes. Thus, engineering cells to better navigate through 3D microenvironments could be part of an effective strategy to enhance efficacy of immune therapeutics.
| S-EPMC8121808 | biostudies-literature
Project description:The development of a multimaterial extrusion bioprinting platform is reported. This platform is capable of depositing multiple coded bioinks in a continuous manner with fast and smooth switching among different reservoirs for rapid fabrication of complex constructs, through digitally controlled extrusion of bioinks from a single printhead consisting of bundled capillaries synergized with programmed movement of the motorized stage.
| S-EPMC5235978 | biostudies-literature
Project description:Additive manufacturing promises to revolutionize manufacturing industries. However, 3D printing of novel build materials is currently limited by constraints inherent to printer designs. In this work, a bench-top powder melt extrusion (PME) 3D printer head was designed and fabricated to print parts directly from powder-based materials rather than filament. The final design of the PME printer head evolved from the Rich Rap Universal Pellet Extruder (RRUPE) design and was realized through an iterative approach. The PME printer was made possible by modifications to the funnel shape, pressure applied to the extrudate by the auger, and hot end structure. Through comparison of parts printed with the PME printer with those from a commercially available fused filament fabrication (FFF) 3D printer using common thermoplastics poly(lactide) (PLA), high impact poly (styrene) (HIPS), and acrylonitrile butadiene styrene (ABS) powders (< 1 mm in diameter), evaluation of the printer performance was performed. For each build material, the PME printed objects show comparable viscoelastic properties by dynamic mechanical analysis (DMA) to those of the FFF objects. However, due to a significant difference in printer resolution between PME (X-Y resolution of 0.8 mm and a Z-layer height calibrated to 0.1 mm) and FFF (X-Y resolution of 0.4 mm and a Z-layer height of 0.18 mm), as well as, an inherently more inconsistent feed of build material for PME than FFF, the resulting print quality, determined by a dimensional analysis and surface roughness comparisons, of the PME printed objects was lower than that of the FFF printed parts based on the print layer uniformity and structure. Further, due to the poorer print resolution and inherent inconsistent build material feed of the PME, the bulk tensile strength and Young's moduli of the objects printed by PME were lower and more inconsistent (49.2 ± 10.7 MPa and 1620 ± 375 MPa, respectively) than those of FFF printed objects (57.7 ± 2.31 MPa and 2160 ± 179 MPa, respectively). Nevertheless, PME print methods promise an opportunity to provide a platform on which it is possible to rapidly prototype a myriad of thermoplastic materials for 3D printing.
| S-EPMC8074846 | biostudies-literature