Precise, high-throughput production of multicellular spheroids with a bespoke 3D bioprinter

Utama, Robert H.; Atapattu, Lakmali; O’Mahony, Aidan P.; Fife, Christopher M.; Baek, Je Hyun; Allard, Théophile; O’Mahony, Kieran J.; Ribeiro, Julio; Gaus, Katharina; Kavallaris, Maria; Gooding, J. Justin

doi:10.1101/2020.04.06.028548

“…Glioblastoma tumor model, [329] neuronal tissue, [330] brain tissue model, [331] contractile smooth muscle tissue [332] SUNP BIOMAKER SunP Biotech International (USA) Extrusion Cancer model, [333] liver [334] T&R Biofab T&R Biofab (South Korea) Extrusion Pre-vascularized cardiac patch, [23] superficial knee cartilage and subchondral bone, [335] organoid-manufacturing [336] WeBio WeBio (Argentina) Extrusion Cartilage regeneration [337] Autodrop Microdrop Inkjet RASTRUM Inventia (Australia) Inkjet Production of multi-cellular spheroids [338] CellJet Digilab (USA) Inkjet Bioprinting of human MSCs [339] Hp D300e Hewlett-Packard (USA) Inkjet Skin model [340] Cellbricks Cellbricks GmbH (Germany) Vat-photopolymerization Liver model [182] NGB-R Poietis (France) LIFT MSC-patterning, [341] skin model Precise Bio Precise Bio (USA) LIFT Ophthalmology [342] Regenova, S-PIKE Cyfuse Biomedical (Japan) Scaffold-free bioprinting Liver tissue model, [343] glioblastoma invasion model [344] and tendon parts, respectively. The bioprinted MTU comprised of both soft and stiff sides, similar to the real tissue.…”

Section: Rx1mentioning

confidence: 99%

Emerging Technologies in Multi‐Material Bioprinting

Ravanbakhsh

¹

,

Karamzadeh

²

,

Bao

³

et al. 2021

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organ-specific tissues, [14] patient-specific grafts, [15] tissue model engineering, [16] and drug screening. [17] The most frequently used technologies for 3D bioprinting include nozzle-based and laser/light-based techniques. Extrusion [18] and inkjet [19] are arguably the most common modalities of nozzle-based bioprinting at the moment, while laser-induced forward transfer (LIFT) [20] and vat-photopolymerization [21] are the two frequently used for laser/lightbased 3D bioprinting. [22] Extrusion bioprinting involves fabrication on a bioprinting platform using bioinks extruded from one or several nozzle(s) (Figure 1A). The extrusion process may be pressure-controlled, with the bioink entrained by means of pneumatic actuation, or flow rate-controlled with the bioink forced by mechanical impulses through syringes. [23] Solidification of the bioprinted structures as they are delivered is obtained through physical, chemical, or photo-crosslinking. [24] Extrusion bioprinting is relatively inexpensive, straightforward, and convenient. It has been embodied, for example, within handheld and portable devices. [25][26][27][28][29] But such convenience must be traded off against significant challenges. Nozzle extrusion necessarily entails a high level of shear stress near the fluidic channel walls. Excessive shear, particularly in high-viscosity bioinks, jeopardizes cell viability. [30] High-resolution bioprinting often requires small-diameter nozzles. The greater shear required imposes a limit on bioink flow rate and throughput. This problem may be addressed through the use of printable shearthinning biomaterials, [31] for which the viscosity decreases under shear stress. Yet, shear-thinning bioink materials that meet all design requirements are not always available. Further notable challenges of extrusion methods include difficulties in finding stable in situ crosslinking methods for nonshear-thinning bioinks, as well as low printing resolution [32] in relation to other methods, and complications in the fabrication of freestanding constructs. [33,34] These shortcomings have spurred many enhancements, notably co-axial/core-shell bioprinting, [35] described in Section 3.1.3, and embedded bioprinting, [36] in Section 3.1.4.Inkjet bioprinting (Figure 1B), similar to home/office inkjet printing, delivers small droplets of bioink to a substrate and can produce high-resolution voxelated constructs. [37] Unlike the extrusion method, where shear-thinning bioinks with a broad range of viscosities can be used, inkjet bioprinters are mainly designed to work with low-viscosity bioinks. [38] The deposition of droplets/voxels is controlled either by thermal, piezoelectric, Bioprinting, within the emerging field of biofabrication, aims at the fabrication of functional biomimetic constructs. Different 3D bioprinting techniques have been adapted to bioprint cell-laden bioinks. However, single-material bioprinting techniques oftentimes fail to reproduce the complex compositions and diversity of native tissues. Multi-material bioprinting as...

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Alginate-gelatin bioink for bioprinting of hela spheroids in alginate-gelatin hexagon shaped scaffolds

Othman

¹

,

Soon

²

,

Ling

³

et al. 2020

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Effect of 3D-bioprinted droplet impact dynamics on a pre-printed soft hydrogel matrix

Chen

¹

,

O’Mahony²,

Barber

³

2023

Exp Fluids

2

0

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Abstract3D droplet-based bioprinting technology is an innovative and time-saving method to precisely generate cell laden 3D structures for multiple clinical and research applications. It is important that the printed droplet must impact so as to leave a single isolated drop, for high printing resolution and accuracy. Therefore, understanding the criteria that control spreading behaviour and prevent droplet splashing is of great importance in optimizing the printing performance. In this experimental work, the physics of the impact of bioink droplets on a pre-printed soft hydrogel matrix was investigated. The droplet size, velocity, and input cell density were varied to generate a range of droplet impact behaviours. It has been shown that the soft substrate inhibited the droplet spreading after impact. The deposition/splashing boundary on the dry/wet flat surface (Nunclon $$^{\text{TM}}$$ TM Delta surface-treated plastic) was defined by $$K = {\text{We}}^{0.5} {\text{Re}} ^{0.25} = 86.19$$ K = We 0.5 Re 0.25 = 86.19 . The splashing threshold on the soft hydrogel matrix was defined by $$K = 44.78$$ K = 44.78 and $$L = {\text{We}} {\text{Re}}^{-0.4} = 13.85$$ L = WeRe - 0.4 = 13.85 for both blank and cell-laden inks on dry/wet soft substrates. Beyond this threshold, the printed droplet volume will be much lower than the expected volume due to splashing, leading to poor printing performance. The absolute splashing threshold on the dry/wet soft hydrogel matrix was defined by $$K = 73.41$$ K = 73.41 and $$L = 27.36$$ L = 27.36 . The printed 3D cell-laden structures were also presented to illustrate how the impact behaviours influence the possible printing fidelity and structure integrity.

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Precise, high-throughput production of multicellular spheroids with a bespoke 3D bioprinter

Cited by 6 publications

References 35 publications

Emerging Technologies in Multi‐Material Bioprinting

Emerging Technologies in Multi‐Material Bioprinting

Alginate-gelatin bioink for bioprinting of hela spheroids in alginate-gelatin hexagon shaped scaffolds

Effect of 3D-bioprinted droplet impact dynamics on a pre-printed soft hydrogel matrix

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