Accessible bioprinting: adaptation of a low-cost 3D-printer for precise cell placement and stem cell differentiation

Abstract

The precision and repeatability offered by computer-aided design and computer-numerically controlled techniques in biofabrication processes is quickly becoming an industry standard. However, many hurdles still exist before these techniques can be used in research laboratories for cellular and molecular biology applications. Extrusion-based bioprinting systems have been characterized by high development costs, injector clogging, difficulty achieving small cell number deposits, decreased cell viability, and altered cell function post-printing. To circumvent the high-price barrier to entry of conventional bioprinters, we designed and 3D printed components for the adaptation of an inexpensive 'off-the-shelf' commercially available 3D printer. We also demonstrate via goal based computer simulations that the needle geometries of conventional commercially standardized, 'luer-lock' syringe-needle systems cause many of the issues plaguing conventional bioprinters. To address these performance limitations we optimized flow within several microneedle geometries, which revealed a short tapered injector design with minimal cylindrical needle length was ideal to minimize cell strain and accretion. We then experimentally quantified these geometries using pulled glass microcapillary pipettes and our modified, low-cost 3D printer. This systems performance validated our models exhibiting: reduced clogging, single cell print resolution, and maintenance of cell viability without the use of a sacrificial vehicle. Using this system we show the successful printing of human induced pluripotent stem cells (hiPSCs) into Geltrex and note their retention of a pluripotent state 7 d post printing. We also show embryoid body differentiation of hiPSC by injection into differentiation conducive environments, wherein we observed continuous growth, emergence of various evaginations, and post-printing gene expression indicative of the presence of all three germ layers. These data demonstrate an accessible open-source 3D bioprinter capable of serving the needs of any laboratory interested in 3D cellular interactions and tissue engineering.

Consistent and reproducible cultures of large-scale 3D mammary epithelial structures using an accessible bioprinting platform

“…However, layer-by-layer methods for 3D cell printing frequently require extrusion nozzles near 300 μm and layer thicknesses of 500 μm, which severely limit the ability to control aspects of the microenvironment at the single-cell scale. We recently described the adaptation of an off-the-shelf 3D printer for the purposes of bioprinting cells within precast 3D substrates [ 26 ]. This system uses pulled glass microneedles, which can be designed with tip diameters ranging from 10 to 100 μm, allowing more accurate cell placement down to the single-cell level.…”

Plant-Derived Biomaterials: A Review of 3D Bioprinting and Biomedical Applications

“…Bioprinting technology is becoming an increasingly available and affordable technology to support tissue engineering research, with many research groups now opting to customize their own in-house devices owing to the widespread availability of open source software and firmware (Reid et al, 2016). A major limitation of bioprinting for biological constructs is the choice of a suitable bioink: one not only biologically conducive of cell growth but in possession of the necessary rheological properties such as viscosity, shear thinning, and crosslinking to optimize post-printing fidelity (Jessop et al, 2017;Kyle et al, 2017).…”

The Biomaker: an entry‐level bioprinting device for biotechnological applications

“…To lower the hurdle for scientists to establish 3D printing technologies in their laboratories entry‐level solutions with a user‐friendly interface are required. To date, only a few studies are available that provide a possibility for a more easy entry to bioprinting by the modification of commercially available non‐bioprinters . These studies present printers specially adapted for the application in the field of TE.…”

“…To date, only a few studies are available that provide a possibility for a more easy entry to bioprinting by the modification of commercially available non-bioprinters. [45][46][47] These studies present printers specially adapted for the application in the field of TE. The particular focus of these works was establishing cell-friendly printing conditions and the preservation of the viability of the printed cells.…”