A gel-free 3D microfluidic cell culture system
Abstract
3D microfluidic cell culture systems offer a biologically relevant model to conduct micro-scale mammalian cell-based research and applications. Various natural and synthetic hydrogels have been successfully incorporated into microfluidic systems to support mammalian cells in 3D. However, embedment of cells in hydrogels introduces operational complexity, potentially hinders mass transfer, and is not suitable for establishing cell-dense, ECM-poor constructs. We present here a gel-free method for seeding and culturing mammalian cells three-dimensionally in a microfluidic channel. A combination of transient inter-cellular polymeric linker and micro-fabricated pillar arrays was used for the in situ formation and immobilization of 3D multi-cellular aggregates in a microfluidic channel. 3D cellular constructs formed this way are relieved of hydrogel embedment for cellular support. Two mammalian cell lines (A549 and C3A) and a primary mammalian cell (bone marrow mesenchymal stem cells) were cultured in the gel-free 3D microfluidic cell culture system. The cells displayed 3D cellular morphology, cellular functions and differentiation capability, affirming the versatility of the system as a 3D cell perfusion culture platform for anchorage-dependent mammalian cells.
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Cells grown in 3D culture can be incorporated into a biosensor either by direct attachment onto a biotic or abiotic substrate surface or by indirect attachment via entrapment in a biocompatible biopolymer.
“Cells grown in 3D culture can be incorporated into a biosensor either by direct attachment onto a biotic or abiotic substrate surface or by indirect attachment via entrapment in a biocompatible biopolymer. 83,84”
From Three-dimensional cell culture systems and their applications in drug discovery and cell-based biosensors (2014) · cites this paper as [84]
The wide variety of commercially available hydrogels promotes facile selection of one or more suitable matrices for each cell line and/or biosensor application and helps overcome disadvantageous features of some polymer matrices.
“The wide variety of commercially available hydrogels promotes facile selection of one or more suitable matrices for each cell line and/or biosensor application and helps overcome the disadvantageous features of some polymer matrices, such as variable compositions and properties (animal-based hydrogels) and requisite cytotoxic pretreatment steps (some synthetic hydrogels). 84”
From Three-dimensional cell culture systems and their applications in drug discovery and cell-based biosensors (2014) · cites this paper as [84]
These 3D cell aggregates preserved cell viability and 3D morphology but exhibited greater cell function compared to a similar 2D monolayer.
“These 3D cell aggregates not only preserved cell viability and 3D morphology but also exhibited greater cell function when compared to similar 2D monolayer. 84”
From Three-dimensional cell culture systems and their applications in drug discovery and cell-based biosensors (2014) · cites this paper as [84]
Ong et al. produced a gel-free 3D system using two carcinoma cell lines and a primary bone marrow mesenchymal stem line.
“Ong et al . produced the gel-free 3D system using two carcinoma cell lines and a primary bone marrow mesenchymal stem line.”
From Three-dimensional cell culture systems and their applications in drug discovery and cell-based biosensors (2014) · cites this paper as [84]
An alternative to the hydrogel matrix for cell lines that reach confluence quickly or naturally have little to no extracellular matrix is a gel-free system that uses an intercellular polymeric linker to create 3D cellular aggregates supported by neighboring cells.
“An alternative to the hydrogel matrix for cell lines that reach confluence quickly or that naturally have little to no extracellular matrix is a gel-free system that uses an intercellular polymeric linker to create 3D cellular aggregates supported by neighboring cells. 84”
From Three-dimensional cell culture systems and their applications in drug discovery and cell-based biosensors (2014) · cites this paper as [84]
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- Authors
- Ong, S.-M., Zhang, C., Toh, Y.-C., Kim, S. H., Foo, H. L., Tan, C. H., van Noort, D., Park, S., & Yu, H.
- Affiliations
- Agency for Science, Technology and Research (SG)
- National University of Singapore (SG)
- Ewha Womans University (KR)
- Venue
- Biomaterials
- Publisher
- Elsevier BV
- Published
- 2008-08
- Volume
- 29
- Issue
- 22
- Pages
- 3237-3244
- ISSN
- 0142-9612 (ISSN-L)
- OpenAlex
- W2124775838
- Cited by
- 172
- References
- 38
- Citations by year
- 2024: 3 · 2023: 4 · 2022: 7 · 2021: 5 · 2020: 8 · 2019: 12 · 2018: 16 · 2017: 11 · 2016: 10 · 2015: 8 · 2014: 15 · 2013: 16 · 2012: 15
- FWCI
- 9.28
- Type
- journal article
- Language
- en
- Open access
- closed
- Locations
- Biomaterials (publishedVersion)
- PubMed (publishedVersion)
- National University of Singapore (submittedVersion)
- Funders
- National University of Singapore
- Ewha Womans University
- National Medical Research Council
- Biomedical Research Council
- Topics
- Physical Sciences › Engineering › Biomedical Engineering › 3D Printing in Biomedical Research
- Physical Sciences › Engineering › Biomedical Engineering › Microfluidic and Capillary Electrophoresis Applications
- Physical Sciences › Engineering › Biomedical Engineering › Microfluidic and Bio-sensing Technologies
- Keywords
- 3D cell culture, microfluidic cell culture, cell aggregation, pillar arrays, cell immobilization, A549 cells, C3A cells, bone marrow mesenchymal stem cells, adherent cells, cell differentiation
- MeSH terms
- Actins, Animals, Bone Marrow Cells, Cell Aggregation, Cell Line, Cell Survival, Cells, Cultured, Humans, Microscopy, Electron, Scanning, Models, Biological, Cell Culture Techniques, Hydrogel, Polyethylene Glycol Dimethacrylate, Microfluidics, Rats, Mesenchymal Stem Cells
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