An optimised low-cost and highly reproducible microfabrication pipeline to generate 3D-printed customised cell culture devices with complex designs

An optimised low-cost and highly reproducible microfabrication pipeline to generate 3D-printed customised cell culture devices with complex designs is reported. The pipeline uses low-cost, high-resolution desktop resin 3D printers in combination with PDMS soft lithography to generate features from μm to cm scale. The pipeline is tailored for biomedical and biological wet labs with limited expertise in microfabrication and will enable them to generate a wide variety of customisable devices for cell culture and tissue engineering in an easy, fast, reproducible, and cost-effective way. The methodology can be easily adopted by any wet lab, irrespective of prior expertise or resource availability, and will enable the wide adoption of tailored microfabricated devices across many fields of biology.

Introduction

Cell culture devices, such as microwells and microfluidic chips, are designed to increase the complexity of cell-based models while retaining control over culture conditions and have become indispensable platforms for biological systems modelling. Whilst their application in biological projects is increasing exponentially, biological and biomedical labs tend more often to rely on already made devices. Commercially developed devices are available for a variety of applications but are often costly and, importantly, lack the potential for customization by each individual lab. Recent important advancements in bioengineering and microfabrication have aimed to solve these issues, and taking advantage of low-cost, high-resolution methods, we have developed an optimised a low-cost and highly reproducible microfabrication pipeline, thought specifically for biomedical and biological wet labs with not prior experience in the field, which will enable them to generate a wide variety of customisable devices for cell culture and tissue engineering with features that vary from 20/50 μm to several centimetres using 3D vat polymerisation. We present a detailed guide on how to use this pipeline, intended for biological and biomedical wet labs, together with the necessary information and context on the techniques involved across the fields of bioengineering and microfabrication and several application examples.

Results

We provide a ready-to-go pipeline for the efficient treatment of resin-based 3D-printed constructs for PDMS curing, using a combination of polymerisation steps, washes, and surface treatments. Together with the extensive characterisation of the fabrication pipeline, we show the utilisation of this system to a variety of applications and use cases relevant to biological experiments, ranging from micro topographies for cell alignments to complex multipart hydrogel culturing systems.

Conclusion

This methodology can be easily adopted by any wet lab, irrespective of prior expertise or resource availability and will enable the wide adoption of tailored microfabricated devices across many fields of biology. 

Availability: All data and procedures are available within the paper and additional information and designs are available as part of this publication and associated files, as well as available to download from the corresponding GitHub repository (https://github.com/SerioLab/SOL3D).

Supplementary Information: Find the supplementary information and figure data directly appended to this file.

Funding: The Serio lab acknowledge support of the UK Biotechnology and Biological Sciences Research Council (BBSRC) [BB/T014318/1] [BB/W006561/1] and of the Dementia Research Institute (UKDRI). F.S.T and A.S. are part of the Horizon Europe "MAGIC" consortium (101080690; www.magic-horizon.eu); this work is funded by UK Research and Innovation (UKRI) under the UK government's Horizon Europe funding guarantee grant numbers 10080927, 10079726, 10082354 and 10078461. Work in the Tedesco lab was also supported by the European Research Council (759108), AFM-Telethon (21687), BBSRC (BB/M009513/1), CureCMD (576031), Muscular Dystrophy UK and the NIHR (the views expressed are those of the authors and not necessarily those of the National Health Service, the NIHR, or the Department of Health). This research was funded in whole, or in part, by the Wellcome Trust. 

Competing interests: The authors have declared that no competing interests exist.

Abbreviations: EB, embryoid body; ECM, extra cellular matrix; GS, goat serum; ICC, immunocytochemistry; IPA, isopropanol; iPSC, induced pluripotent stem cell; MN, motor neuron; MNP, motor neuron progenitor; PDMS, polydimethylsiloxane; SOL3D, softlithography on 3D vat polymerised moulds