Rapid Magnetic 3D Printing of Cellular Structures with MCF-7 Cell Inks

Mishriki, Sarah; Fattah, Abdel Rahman Abdel; Kammann, Tobias; Sahu, Rakesh P.; Geng, Fei; Puri, Ishwar K.

doi:10.34133/2019/9854593

Cited by 18 publications

(17 citation statements)

References 42 publications

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“…Assess the Susceptibility of 3D Cellular Structures to Gd-DTPA. As previously demonstrated, the printing of 3D cellular structures with magnetic assistance is facilitated by adding 25 mM Gd-DTPA to the cell culture medium [23][24][25]. During the formation of a 3D MCF-7 cellular structure, the limiting exposure time to Gd-DTPA was established by evaluating the relative changes in cell viability.…”

Section: Presence Of Gd 3+ In Monotypic 3d Cellular Structures Tomentioning

confidence: 99%

“…Cell viability was not significantly affected by exposure to 25 mM Gd-DTPA for 24 hours. The 3D structure was printed within 6 hours for specific conditions that include seeding cell density, magnet dimension, and well size [23].…”

Section: Presence Of Gd 3+ In Monotypic 3d Cellular Structures Tomentioning

confidence: 99%

“…Printing 3D cellular structures with magnetic assistance requires (1) a suspension of cells in a liquid medium, (2) a magnetic susceptibility difference between the cells (diamagnetic) and the medium (paramagnetic), and (3) an applied inhomogeneous magnetic field. After the 3D structures are printed, the magnetic field can be removed and the paramagnetic medium replaced with a usual cell medium [23]. However, if the paramagnetic cell medium is replaced with a Gd-free cell medium prior to sufficient intercellular interactions occurring, the cell agglomerate is readily disturbed and the morphology of the resulting 3D structure distorted.…”

Section: Formation Of Monotypic and Coculture 3d Cellularmentioning

confidence: 99%

“…Magnetic printing is an engineering solution to create reproducible 3D cellular structures that can be used for in vitro cellular studies [ 23 – 28 ]. Here, using a unique bottom-up approach, 3D cellular assemblies can be formed by exploiting the magnetic properties of cells.…”

Section: Introductionmentioning

confidence: 99%

“…Within the appropriate exposure limits, Gd-DTPA (a magnetic resonance imaging (MRI) contrast agent (CA)) is potentially nontoxic. The addition of Gd-DTPA establishes a magnetic susceptibility difference between the diamagnetic cells and their surrounding paramagnetic liquid media and has been shown to effectively facilitate the displacement of cells in the presence of a magnetic field [ 23 , 24 , 26 , 27 ]. In an inhomogeneous magnetic field, the suspended cells are displaced towards regions of lowest magnetic field strength to form a single 3D cellular cluster in a contactless, label-free manner within hours ( Figure 1 and Video S1 ).…”

Section: Introductionmentioning

confidence: 99%

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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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Section: Presence Of Gd 3+ In Monotypic 3d Cellular Structures Tomentioning

confidence: 99%

Section: Presence Of Gd 3+ In Monotypic 3d Cellular Structures Tomentioning

confidence: 99%

Section: Formation Of Monotypic and Coculture 3d Cellularmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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

et al. 2020

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Levitational 3D Bioassembly and Density‐Based Spatial Coding of Levitoids

Moncal

Yaman

Durmus

2022

Adv Funct Materials

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Self‐assembly of cells into functional bioarchitectures with microscale spatial topographies are prevalent in nature. Despite the advances to recapitulate the native‐like microenvironment in vitro,fabrication of soft living materials with a deterministic control on the composition, functionality, and geometry, is still challenging. Here, a versatile, paramagnetically tunable, levitational biofabrication technique within a ring‐magnet system, RM‐LEV, is developed to assemble single cells or heterogeneous multicellular living architectures in a scaffold‐free manner. The self‐assembly and contactless biofabrication of multiple cell types into 3D multicellular “levitospheres” is demonstrated, where cellular positions are guided by levitation and density, simultaneously. Inherent density and diamagnetism of different cell types are leveraged to manipulate the geometry and self‐assembly of levitospheres into larger complex 3D structures, termed as “levitoids”. This approach is further applied to manipulate, position, and culture levitoids within a 3D hydrogel matrix under levitation, which preserves their viability and functionality. Thus, this study provides a foundation to create spatially heterogeneous 3D cellular assemblies with density‐coded bio‐architectures—which is not previously realized using magnetic levitation. This density‐based coding approach can be beneficial to study the cross‐talk between different cell types within spheroids and organoids, and be broadly applied to 3D bioprinting, tissue engineering, cancer, and neuroscience research.

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Label‐Free Cell Migration Assay Using Magnetic Exclusion

Gupta

Dabaghi

et al. 2021

Adv Materials Technologies

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biophysical cues. [1,2] Signaling molecules regulate cell migration under normal physiological conditions by controlling cell adhesion and cytoskeleton organization. [1,3,4] Conversely, pathological conditions such as metastatic cancer, vascular disease, and chronic inflammation are associated with unregulated cell migration, contributing to disease progression. [4,5] The wound healing assay is a commonly used in vitro method to study cell migration, cell interactions, and the underlying mechanisms. [6][7][8][9][10] The conventional "scratch" assay creates a cellfree area or scratch in a cell monolayer using a pipette tip or a pin tool. [10,11] Cells from intact surrounding regions migrate and repopulate the cell-free region over time. The migration rate is measured by observing the decrease in the cellfree area at regular intervals. While the scratch assay is simple and inexpensive, it requires 24-48 h to form a cell monolayer. [11] Moreover, by physically scratching the cell layer with a pipette tip or microneedle, cell-free regions of inconsistent shapes and sizes are created. [10,12,13] This process also damages the underlying plastic surface or extracellular matrix (ECM) substrate, which can significantly affect cell migration. [10,12,13] The electric cell-substrate impedance sensing (ECIS) technique, on the other hand, produces consistent cell-free regions without mechanically disrupting the cell layer. [10,12] In ECIS, a confluent cell layer is formed over a small gold film electrode placed at the bottom of a tissue culture well. Electric pulses are applied to induce cell death at the electrode surface, which creates a well-defined region devoid of cells. [14] However, dead cell fragments can remain on top of the electrode and interfere with cell migration. [12,15] Furthermore, the electric pulses can potentially injure the cells surrounding the electrode, thereby affecting their migration. [10] In contrast to the scratch assay and ECIS, the physical exclusion technique creates cell-free areas of reproducible sizes and shapes with minimal cellular damage. [12,16] This method employs physical barriers, such as plastic inserts or silicone stoppers, to prevent cells from settling in a predefined area. However, it requires 12-20 h to form confluent cellular regions around the barrier. [16,17] Commercial inserts are also expensive and unsuitable for multiple uses. [16] Magnetic levitation is another in vitro technique used to form cell clusters in a migration assay. [18] This method is based on the cellular uptake of biopolymer-encapsulated magnetic In vitro wound healing assays are widely used to investigate cell migration during various physiologic and pathologic processes. However, traditional scratch-based assays produce cell-free areas that are not reproducible, whereas the alternate insert-based exclusion method is expensive and timeconsuming. Here, a rapid, label-free, insert-free magnetic exclusion technique, where magnetic fields are used to create cell-free areas is described. Suspensions of diamagnet...

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Rapid Magnetic 3D Printing of Cellular Structures with MCF-7 Cell Inks

Cited by 18 publications

References 42 publications

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

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

Levitational 3D Bioassembly and Density‐Based Spatial Coding of Levitoids

Label‐Free Cell Migration Assay Using Magnetic Exclusion

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