Low-cost, versatile, and highly reproducible microfabrication pipeline to generate 3D-printed customised cell culture devices with complex designs

Cathleen Hagemann; Matthew C D Bailey; Eugenia Carraro; Ksenia S Stankevich; Valentina Maria Lionello; Noreen Khokhar; Pacharaporn Suklai; Carmen Moreno-Gonzalez; Kelly O'Toole; George Konstantinou; Christina L Dix; Sudeep Joshi; Eleonora Giagnorio; Mads S Bergholt; Christopher D Spicer; Albane Imbert; Francesco Saverio Tedesco; Andrea Serio

doi:10.1371/journal.pbio.3002503

Low-cost, versatile, and highly reproducible microfabrication pipeline to generate 3D-printed customised cell culture devices with complex designs

Cathleen Hagemann, Matthew C D Bailey, Eugenia Carraro, Ksenia S Stankevich, Valentina Maria Lionello, Noreen Khokhar, Pacharaporn Suklai, Carmen Moreno-Gonzalez, Kelly O'Toole, George Konstantinou, Christina L Dix, Sudeep Joshi, Eleonora Giagnorio, Mads S Bergholt, Christopher D Spicer, Albane Imbert, Francesco Saverio Tedesco, Andrea Serio

Research output: Contribution to journal › Article › peer-review

2 Citations (Scopus)

Abstract

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. From microtopography, microwells, plating devices, and microfluidic systems to larger constructs such as live imaging chamber slides, a wide variety of culture devices with different geometries have become indispensable in biology laboratories. However, while their application in biological projects is increasing exponentially, due to a combination of the techniques, equipment and tools required for their manufacture, and the expertise necessary, biological and biomedical labs tend more often to rely on already made devices. Indeed, commercially developed devices are available for a variety of applications but are often costly and, importantly, lack the potential for customisation by each individual lab. The last point is quite crucial, as often experiments in wet labs are adapted to whichever design is already available rather than designing and fabricating custom systems that perfectly fit the biological question. This combination of factors still restricts widespread application of microfabricated custom devices in most biological wet labs. Capitalising on recent advances in bioengineering and microfabrication aimed at solving these issues, and taking advantage of low-cost, high-resolution desktop resin 3D printers combined with PDMS soft lithography, we have developed an optimised a low-cost and highly reproducible microfabrication pipeline. This is 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 in an easy, fast reproducible way for a fraction of the cost of conventional microfabrication or commercial alternatives. This protocol is designed specifically to be a resource for biological labs with limited expertise in those techniques and enables the manufacture of complex devices across the μm to cm scale. 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. 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.

Original language	English
Pages (from-to)	e3002503
Journal	PLoS Biology
Volume	22
Issue number	3
DOIs	https://doi.org/10.1371/journal.pbio.3002503
Publication status	Published - 13 Mar 2024

Keywords

Microtechnology
Cell Culture Techniques
Microfluidics/methods
Printing, Three-Dimensional
Lab-On-A-Chip Devices

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

Access to Document

10.1371/journal.pbio.3002503

Cite this

Hagemann, C., Bailey, M. C. D., Carraro, E., Stankevich, K. S., Lionello, V. M., Khokhar, N., Suklai, P., Moreno-Gonzalez, C., O'Toole, K., Konstantinou, G., Dix, C. L., Joshi, S., Giagnorio, E., Bergholt, M. S., Spicer, C. D., Imbert, A., Tedesco, F. S., & Serio, A. (2024). Low-cost, versatile, and highly reproducible microfabrication pipeline to generate 3D-printed customised cell culture devices with complex designs. PLoS Biology, 22(3), e3002503. https://doi.org/10.1371/journal.pbio.3002503

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title = "Low-cost, versatile, and highly reproducible microfabrication pipeline to generate 3D-printed customised cell culture devices with complex designs",

abstract = "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. From microtopography, microwells, plating devices, and microfluidic systems to larger constructs such as live imaging chamber slides, a wide variety of culture devices with different geometries have become indispensable in biology laboratories. However, while their application in biological projects is increasing exponentially, due to a combination of the techniques, equipment and tools required for their manufacture, and the expertise necessary, biological and biomedical labs tend more often to rely on already made devices. Indeed, commercially developed devices are available for a variety of applications but are often costly and, importantly, lack the potential for customisation by each individual lab. The last point is quite crucial, as often experiments in wet labs are adapted to whichever design is already available rather than designing and fabricating custom systems that perfectly fit the biological question. This combination of factors still restricts widespread application of microfabricated custom devices in most biological wet labs. Capitalising on recent advances in bioengineering and microfabrication aimed at solving these issues, and taking advantage of low-cost, high-resolution desktop resin 3D printers combined with PDMS soft lithography, we have developed an optimised a low-cost and highly reproducible microfabrication pipeline. This is 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 in an easy, fast reproducible way for a fraction of the cost of conventional microfabrication or commercial alternatives. This protocol is designed specifically to be a resource for biological labs with limited expertise in those techniques and enables the manufacture of complex devices across the μm to cm scale. 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. 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.",

keywords = "Microtechnology, Cell Culture Techniques, Microfluidics/methods, Printing, Three-Dimensional, Lab-On-A-Chip Devices",

author = "Cathleen Hagemann and Bailey, {Matthew C D} and Eugenia Carraro and Stankevich, {Ksenia S} and Lionello, {Valentina Maria} and Noreen Khokhar and Pacharaporn Suklai and Carmen Moreno-Gonzalez and Kelly O'Toole and George Konstantinou and Dix, {Christina L} and Sudeep Joshi and Eleonora Giagnorio and Bergholt, {Mads S} and Spicer, {Christopher D} and Albane Imbert and Tedesco, {Francesco Saverio} and Andrea Serio",

note = "Copyright: {\textcopyright} 2024 Hagemann et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.",

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Hagemann, C , Bailey, MCD , Carraro, E, Stankevich, KS, Lionello, VM, Khokhar, N, Suklai, P , Moreno-Gonzalez, C , O'Toole, K, Konstantinou, G, Dix, CL, Joshi, S, Giagnorio, E, Bergholt, MS, Spicer, CD, Imbert, A, Tedesco, FS & Serio, A 2024, 'Low-cost, versatile, and highly reproducible microfabrication pipeline to generate 3D-printed customised cell culture devices with complex designs', PLoS Biology, vol. 22, no. 3, pp. e3002503. https://doi.org/10.1371/journal.pbio.3002503

TY - JOUR

T1 - Low-cost, versatile, and highly reproducible microfabrication pipeline to generate 3D-printed customised cell culture devices with complex designs

AU - Hagemann, Cathleen

AU - Bailey, Matthew C D

AU - Carraro, Eugenia

AU - Stankevich, Ksenia S

AU - Lionello, Valentina Maria

AU - Khokhar, Noreen

AU - Suklai, Pacharaporn

AU - Moreno-Gonzalez, Carmen

AU - O'Toole, Kelly

AU - Konstantinou, George

AU - Dix, Christina L

AU - Joshi, Sudeep

AU - Giagnorio, Eleonora

AU - Bergholt, Mads S

AU - Spicer, Christopher D

AU - Imbert, Albane

AU - Tedesco, Francesco Saverio

AU - Serio, Andrea

N1 - Copyright: © 2024 Hagemann et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

PY - 2024/3/13

Y1 - 2024/3/13

N2 - 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. From microtopography, microwells, plating devices, and microfluidic systems to larger constructs such as live imaging chamber slides, a wide variety of culture devices with different geometries have become indispensable in biology laboratories. However, while their application in biological projects is increasing exponentially, due to a combination of the techniques, equipment and tools required for their manufacture, and the expertise necessary, biological and biomedical labs tend more often to rely on already made devices. Indeed, commercially developed devices are available for a variety of applications but are often costly and, importantly, lack the potential for customisation by each individual lab. The last point is quite crucial, as often experiments in wet labs are adapted to whichever design is already available rather than designing and fabricating custom systems that perfectly fit the biological question. This combination of factors still restricts widespread application of microfabricated custom devices in most biological wet labs. Capitalising on recent advances in bioengineering and microfabrication aimed at solving these issues, and taking advantage of low-cost, high-resolution desktop resin 3D printers combined with PDMS soft lithography, we have developed an optimised a low-cost and highly reproducible microfabrication pipeline. This is 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 in an easy, fast reproducible way for a fraction of the cost of conventional microfabrication or commercial alternatives. This protocol is designed specifically to be a resource for biological labs with limited expertise in those techniques and enables the manufacture of complex devices across the μm to cm scale. 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. 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.

AB - 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. From microtopography, microwells, plating devices, and microfluidic systems to larger constructs such as live imaging chamber slides, a wide variety of culture devices with different geometries have become indispensable in biology laboratories. However, while their application in biological projects is increasing exponentially, due to a combination of the techniques, equipment and tools required for their manufacture, and the expertise necessary, biological and biomedical labs tend more often to rely on already made devices. Indeed, commercially developed devices are available for a variety of applications but are often costly and, importantly, lack the potential for customisation by each individual lab. The last point is quite crucial, as often experiments in wet labs are adapted to whichever design is already available rather than designing and fabricating custom systems that perfectly fit the biological question. This combination of factors still restricts widespread application of microfabricated custom devices in most biological wet labs. Capitalising on recent advances in bioengineering and microfabrication aimed at solving these issues, and taking advantage of low-cost, high-resolution desktop resin 3D printers combined with PDMS soft lithography, we have developed an optimised a low-cost and highly reproducible microfabrication pipeline. This is 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 in an easy, fast reproducible way for a fraction of the cost of conventional microfabrication or commercial alternatives. This protocol is designed specifically to be a resource for biological labs with limited expertise in those techniques and enables the manufacture of complex devices across the μm to cm scale. 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. 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.

KW - Microtechnology

KW - Cell Culture Techniques

KW - Microfluidics/methods

KW - Printing, Three-Dimensional

KW - Lab-On-A-Chip Devices

U2 - 10.1371/journal.pbio.3002503

DO - 10.1371/journal.pbio.3002503

M3 - Article

C2 - 38478490

SN - 1544-9173

VL - 22

SP - e3002503

JO - PLoS Biology

JF - PLoS Biology

IS - 3

ER -

Low-cost, versatile, and highly reproducible microfabrication pipeline to generate 3D-printed customised cell culture devices with complex designs

Abstract

Keywords

UN SDGs

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