A rapid biofabrication technique for self-assembled collagen-based multicellular and heterogeneous 3D tissue constructs

Shahin‐Shamsabadi, Alireza; Selvaganapathy, P. Ravi

doi:10.1016/j.actbio.2019.05.024

Cited by 23 publications

(29 citation statements)

References 42 publications

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“…The ability to form 3D homogeneous and heterogeneous constructs with control over positioning of cells [ 24 ] using fat and muscle cells allows, for the first time, the study of direct (cell–cell) and indirect (through paracrine activity) interactions between the cell types in 3D, delineate them and understand the mechanisms through which their growth, metabolic activity, and regulation are modulated. Nevertheless, such constructs should have structural integrity despite the morphological changes that will be introduced during the self‐assembly process to form the constructs and the remodeling that happens later on, as well as the respective cell differentiation processes.…”

Section: Resultsmentioning

confidence: 99%

“…Homogeneous and heterogeneous 3D tissue constructs composed of adipocytes and skeletal muscle cells in bovine collagen type I as ECM were fabricated using a newly developed rapid self‐assembly process. [ 24 ] A poly(lactic acid) master mold shaped in the form of two intersecting cylindrical posts (each of them were 3 mm in diameter and 3 mm in height) with a 0.5 mm overlap was 3D printed using a stereolithography printer (Objet 24 Desktop 3D printer). Polydimethylsiloxane (PDMS) at 10:1 ratio of the polymer to its curing agent was cast onto the master mold to form the microwells ( Figure 1 a) that were used to form the self‐assembled constructs for 3D culture systems.…”

Section: Methodsmentioning

confidence: 99%

“…In this paper, we use a newly developed rapid 3D tissue construct fabrication method as the tool to fabricate co‐culture systems with controlled positioning of cells [ 24 ] and create the first spatially defined, heterogeneous tissue construct composed of adipocyte and skeletal muscle cells. We study effect of media type on the behavior of both of cell types in co‐culture and integrity of the 3D constructs.…”

Section: Introductionmentioning

confidence: 99%

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A 3D Self‐Assembled In Vitro Model to Simulate Direct and Indirect Interactions between Adipocytes and Skeletal Muscle Cells

Shahin‐Shamsabadi

Selvaganapathy

2020

Adv. Biosys.

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The molecular mechanisms of the development and progression of diabetes and obesity involve complex interactions between adipocytes and skeletal muscle cells. Although 2D in‐vitro models are the gold standard for the mechanistic study of such behaviors, they do not recreate the complexity and dynamics of the interactions between the cell types involved. Alternatively, animal models are used but are expensive, difficult to visualize or analyze, are not completely representative of human physiology or genetic background, and have associated ethical considerations. 3D co‐culture systems can be complementary to these approaches. Here, using a newly developed 3D biofabrication method, adipocytes and myoblasts are positioned precisely either in direct physical contact or in close proximity such that the paracrine effects could be systematically studied. Suitable protocols for growth and differentiation of both cells in the co‐culture system is also developed. Cells show more restrained lipid and protein production in 3D systems compared to 2D ones and adipocytes show more lipolysis in indirect contact with myoblasts as response to drug treatment. These findings emphasize importance of physical contact between cells that have been overlooked in co‐culture systems using transwell inserts and can be used in studies for the development of anti‐obesity drugs.

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Section: Resultsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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A 3D Self‐Assembled In Vitro Model to Simulate Direct and Indirect Interactions between Adipocytes and Skeletal Muscle Cells

Shahin‐Shamsabadi

Selvaganapathy

2020

Adv. Biosys.

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“…Although NIH/3T3 cells are of animal origin, their intended use in this study is to evaluate the influence of fibroblasts on the formation of 3D cellular structures for cell types which are difficult to cohere without using additional reagents. Such a use of cell lines from different species has been used in previous coculture models [ 33 , 63 ]. Since human and mouse fibroblasts behave similarly in terms of their ability to produce ECM proteins (such as collagen) [ 64 – 66 ], the conclusions of our study demonstrate the capability of magnetically assisted printing in this coculture model.…”

Section: Discussionmentioning

confidence: 99%

Fibroblasts Accelerate Formation and Improve Reproducibility of 3D Cellular Structures Printed with Magnetic Assistance

et al. 2020

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Fibroblasts (mouse, NIH/3T3) are combined with MDA-MB-231 cells to accelerate the formation and improve the reproducibility of 3D cellular structures printed with magnetic assistance. Fibroblasts and MDA-MB-231 cells are cocultured to produce 12.5 : 87.5, 25 : 75, and 50 : 50 total population mixtures. These mixtures are suspended in a cell medium containing a paramagnetic salt, Gd-DTPA, which increases the magnetic susceptibility of the medium with respect to the cells. A 3D monotypic MDA-MB-231 cellular structure is printed within 24 hours with magnetic assistance, whereas it takes 48 hours to form a similar structure through gravitational settling alone. The maximum projected areas and circularities, and cellular ATP levels of the printed structures are measured for 336 hours. Increasing the relative amounts of the fibroblasts mixed with the MDA-MB-231 cells decreases the time taken to form the structures and improves their reproducibility. Structures produced through gravitational settling have larger maximum projected areas and cellular ATP, but are deemed less reproducible. The distribution of individual cell lines in the cocultured 3D cellular structures shows that printing with magnetic assistance yields 3D cellular structures that resemble in vivo tumors more closely than those formed through gravitational settling. The results validate our hypothesis that (1) fibroblasts act as a “glue” that supports the formation of 3D cellular structures, and (2) the structures are produced more rapidly and with greater reproducibility with magnetically assisted printing than through gravitational settling alone. Printing of 3D cellular structures with magnetic assistance has applications relevant to drug discovery, lab-on-chip devices, and tissue engineering.

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“…Collagenous constructs of cells inside a silicone tubing and anchored to the metallic pins were formed using a process of self-assembly that has been used previously for spheroidal and non-spheroidal structures [ 27 ]. There, a mixture of cells, medium, and neutralized bovine collagen type I were added to a polydimethylsiloxane (PDMS) mold with the specific form that defined the final shape of the construct.…”

Section: Methodsmentioning

confidence: 99%

Tissue-in-a-Tube: three-dimensional in vitro tissue constructs with integrated multimodal environmental stimulation

Shahin‐Shamsabadi

Selvaganapathy

2020

Materials Today Bio

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Three-dimensional (3D) in vitro tissue models are superior to two-dimensional (2D) cell cultures in replicating natural physiological/pathological conditions by recreating the cellular and cell-matrix interactions more faithfully. Nevertheless, current 3D models lack either the rich multicellular environment or fail to provide appropriate biophysical stimuli both of which are required to properly recapitulate the dynamic in vivo microenvironment of tissues and organs. Here, we describe the rapid construction of multicellular, tubular tissue constructs termed Tissue-in-a-Tube using self-assembly process in tubular molds with the ability to incorporate a variety of biophysical stimuli such as electrical field, mechanical deformation, and shear force of the fluid flow. Unlike other approaches, this method is simple, requires only oxygen permeable silicone tubing that molds the tissue construct and thin stainless-steel pins inserted in it to anchor the construct and could be used to provide electrical and mechanical stimuli, simultaneously. The annular region between the tissue construct and the tubing is used for perfusion. Highly stable, macroscale, and robust constructs anchored to the pins form as a result of self-assembly of the extracellular matrix (ECM) and cells in the bioink that is filled into the tubing. We demonstrate patterning of grafts containing cell types in the constructs in axial and radial modes with clear interface and continuity between the layers. Different environmental factors affecting cell behavior such as compactness of the structure and size of the constructs can be controlled through parameters such as initial cell density, ECM content, tubing size, as well as the distance between anchor pins. Using connectors, network of tubing can be assembled to create complex macrostructured tissues (centimeters length) such as fibers that are bifurcated or columns with different axial thicknesses which can then be used as building blocks for biomimetic constructs or tissue regeneration. The method is versatile and compatible with various cell types including endothelial, epithelial, skeletal muscle cells, osteoblast cells, and neuronal cells. As an example, long mature skeletal muscle and neuronal fibers as well as bone constructs were fabricated with cellular alignment dictated by the applied electrical field. The versatility, speed, and low cost of this method is suited for widespread application in tissue engineering and regenerative medicine.

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A rapid biofabrication technique for self-assembled collagen-based multicellular and heterogeneous 3D tissue constructs

Cited by 23 publications

References 42 publications

A 3D Self‐Assembled In Vitro Model to Simulate Direct and Indirect Interactions between Adipocytes and Skeletal Muscle Cells

A 3D Self‐Assembled In Vitro Model to Simulate Direct and Indirect Interactions between Adipocytes and Skeletal Muscle Cells

Fibroblasts Accelerate Formation and Improve Reproducibility of 3D Cellular Structures Printed with Magnetic Assistance

Tissue-in-a-Tube: three-dimensional in vitro tissue constructs with integrated multimodal environmental stimulation

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