skip to main content


Title: Rapid prototyping of cell culture microdevices using parylene-coated 3D prints
Fabrication of microfluidic devices by photolithography generally requires specialized training and access to a cleanroom. As an alternative, 3D printing enables cost-effective fabrication of microdevices with complex features that would be suitable for many biomedical applications. However, commonly used resins are cytotoxic and unsuitable for devices involving cells. Furthermore, 3D prints are generally refractory to elastomer polymerization such that they cannot be used as master molds for fabricating devices from polymers ( e.g. polydimethylsiloxane, or PDMS). Different post-print treatment strategies, such as heat curing, ultraviolet light exposure, and coating with silanes, have been explored to overcome these obstacles, but none have proven universally effective. Here, we show that deposition of a thin layer of parylene, a polymer commonly used for medical device applications, renders 3D prints biocompatible and allows them to be used as master molds for elastomeric device fabrication. When placed in culture dishes containing human neurons, regardless of resin type, uncoated 3D prints leached toxic material to yield complete cell death within 48 hours, whereas cells exhibited uniform viability and healthy morphology out to 21 days if the prints were coated with parylene. Diverse PDMS devices of different shapes and sizes were easily cast from parylene-coated 3D printed molds without any visible defects. As a proof-of-concept, we rapid prototyped and tested different types of PDMS devices, including triple chamber perfusion chips, droplet generators, and microwells. Overall, we suggest that the simplicity and reproducibility of this technique will make it attractive for fabricating traditional microdevices and rapid prototyping new designs. In particular, by minimizing user intervention on the fabrication and post-print treatment steps, our strategy could help make microfluidics more accessible to the biomedical research community.  more » « less
Award ID(s):
2033800
NSF-PAR ID:
10302698
Author(s) / Creator(s):
 ;  ;  ;  ;  ;  ;  ;  ;  ;  ;  ;  ;  ;  ;  
Date Published:
Journal Name:
Lab on a Chip
Volume:
21
Issue:
24
ISSN:
1473-0197
Format(s):
Medium: X
Sponsoring Org:
National Science Foundation
More Like this
  1. Nguyen, Nam-Trung ; Munoz, Rodrigo Alejandro ; Kalinke, Cristiane (Ed.)
    Engineering microfluidic devices relies on the ability to manufacture sub-100 micrometer fluidic channels. Conventional lithographic methods provide high resolution but require costly exposure tools and outsourcing of masks, which extends the turnaround time to several days. The desire to accelerate design/test cycles has motivated the rapid prototyping of microfluidic channels; however, many of these methods (e.g., laser cutters, craft cutters, fused deposition modeling) have feature sizes of several hundred microns or more. In this paper, we describe a 1-day process for fabricating sub-100 µm channels, leveraging a low-cost (USD 600) 8K digital light projection (DLP) 3D resin printer. The soft lithography process includes mold printing, post-treatment, and casting polydimethylsiloxane (PDMS) elastomer. The process can produce microchannels with 44 µm lateral resolution and 25 µm height, posts as small as 400 µm, aspect ratio up to 7, structures with varying z-height, integrated reservoirs for fluidic connections, and a built-in tray for casting. We discuss strategies to obtain reliable structures, prevent mold warpage, facilitate curing and removal of PDMS during molding, and recycle the solvents used in the process. To our knowledge, this is the first low-cost 3D printer that prints extruded structures that can mold sub-100 µm channels, providing a balance between resolution, turnaround time, and cost (~USD 5 for a 2 × 5 × 0.5 cm^3 chip) that will be attractive for many microfluidics labs. 
    more » « less
  2. Abstract

    Polydimethylsiloxane (PDMS) is an inexpensive robust polymer that is commonly used as the fundamental fabrication material for soft‐lithography‐based microfluidic devices. Owing to its versatile material properties, there are some attempts to use PDMS as a porous 3D structure for sensing. However, reliable and easy fabrication has been challenging along with the inherent hydrophobic nature of PDMS hindering its use in biomedical sensing applications. Herein, a cleanroom‐free inexpensive method to create 3D porous PDMS structures, “ExoSponge” and the effective surface modification to functionalize its 3D porous structure is reported. The ability of ExoSponge to recover cancer‐associated extracellular vesicles (EVs) from complex biological samples of up to 10 mL in volume is demonstrated. When compared to ultracentrifugation, the ExoSponge showes a significant increase in cancer EV isolation of more than 210%. Targeted ultra‐high pressure liquid chromatography‐tandem mass spectrometry (LC‐MS/MS) is further employed to profile 70 metabolites in the EVs recovered from the lung cancer patient's plasma. The resulting profiles reveal the potential intraexosomal metabolite biomarker, phenylacetylglutamine (PAG), in non‐small cell lung cancer. The high sensitivity, simple usage, and cost‐effectiveness of the ExoSponge platform creates huge potential for rapid, economical and yet specific isolation of exosomes enabling future diagnostic applications of EVs in cancers.

     
    more » « less
  3. Abstract

    3D printing has become one of the primary fabrication strategies used in biomedical research. Recent efforts have focused on the 3D printing of hydrogels to create structures that better replicate the mechanical properties of biological tissues. These pose a unique challenge, as soft materials are difficult to pattern in three dimensions with high fidelity. Currently, a small number of biologically derived polymers that form hydrogels are frequently reused for 3D printing applications. Thus, there exists a need for novel hydrogels with desirable biological properties that can be used as 3D printable inks. In this work, the printability of multidomain peptides (MDPs), a class of self‐assembling peptides that form a nanofibrous hydrogel at low concentrations, is established. MDPs with different charge functionalities are optimized as distinct inks and are used to create complex 3D structures, including multi‐MDP prints. Additionally, printed MDP constructs are used to demonstrate charge‐dependent differences in cellular behavior in vitro. This work presents the first time that self‐assembling peptides have been used to print layered structures with overhangs and internal porosity. Overall, MDPs are a promising new class of 3D printable inks that are uniquely peptide‐based and rely solely on supramolecular mechanisms for assembly.

     
    more » « less
  4. Recent developments in micro-scale additive manufacturing (AM) have opened new possibilities in state-of-the-art areas, including microelectromechanical systems (MEMS) with intrinsically soft and compliant components. While fabrication with soft materials further complicates micro-scale AM, a soft photocurable polydimethylsiloxane (PDMS) resin, IP-PDMS, has recently entered the market of two-photon polymerization (2PP) AM. To facilitate the development of microdevices with soft components through the application of 2PP technique and IP-PDMS material, this research paper presents a comprehensive material characterization of IP-PDMS. The significance of this study lies in the scarcity of existing research on this material and the thorough investigation of its properties, many of which are reported here for the first time. Particularly, for uncured IP-PDMS resin, this work evaluates a surface tension of 26.7 ± 4.2 mN/m, a contact angle with glass of 11.5 ± 0.6°, spin-coating behavior, a transmittance of more than 90% above 440 nm wavelength, and FTIR with all the properties reported for the first time. For cured IP-PDMS, novel characterizations include a small mechanical creep, a velocity-dependent friction coefficient with glass, a typical dielectric permittivity value of 2.63 ± 0.02, a high dielectric/breakdown strength for 3D-printed elastomers of up to 73.3 ± 13.3 V/µm and typical values for a spin coated elastomer of 85.7 ± 12.4 V/µm, while the measured contact angle with water of 103.7 ± 0.5°, Young’s modulus of 5.96 ± 0.2 MPa, and viscoelastic DMA mechanical characterization are compared with the previously reported values. Friction, permittivity, contact angle with water, and some of the breakdown strength measurements were performed with spin-coated cured IP-PDMS samples. Based on the performed characterization, IP-PDMS shows itself to be a promising material for micro-scale soft MEMS, including microfluidics, storage devices, and micro-scale smart material technologies.

     
    more » « less
  5. Combination of stem cell technology and 3D biofabrication approaches provides physiological similarity to in vivo tissues and the capability of repairing and regenerating damaged human tissues. Mesenchymal stem cells (MSCs) have been widely used for regenerative medicine applications because of their immunosuppressive properties and multipotent potentials. To obtain large amount of high-quality MSCs without patient donation and invasive procedures, we differentiated MSCs from human-induced pluripotent stem cells (hiPSC-MSCs) using serum-free E6 media supplemented with only one growth factor (bFGF) and two small molecules (SB431542 and CHIR99021). The differentiated cells showed a high expression of common MSC-specific surface markers (CD90, CD73, CD105, CD106, CD146, and CD166) and a high potency for osteogenic and chondrogenic differentiation. With these cells, we have been able to manufacture MSC tissue rings with high consistency and robustness in pluronic-coated reusable PDMS devices. The MSC tissue rings were characterized based on inner diameter and outer ring diameter and observed cell-type-dependent tissue contraction induced by cell-matrix interaction. Our approach of simplified hiPSC-MSC differentiation, modular fabrication procedure, and serum-free culture conditions has a great potential for scalable manufacturing of MSC tissue rings for different regenerative medicine applications. 
    more » « less