Abstract:Three-dimensional (3D) cell culture systems have gained increasing interest in drug discovery and tissue engineering due to their evident advantages in providing more physiologically relevant information and more predictive data for in vivo tests. In this review, we discuss the characteristics of 3D cell culture systems in comparison to the two-dimensional (2D) monolayer culture, focusing on cell growth conditions, cell proliferation, population, and gene and protein expression profiles. The innovations and de… Show more
“…[48][49] The Lutolf group has demonstrated the use of 3D hydrogels in a microarray made using automated liquid handling robotics to test many microenvironmental conditions on the regulation of mouse embryonic stem cell self-renewal. 50 This study was a valuable demonstration of the utility of biomaterials in a high-throughput format and it was an important contribution towards using synthetic materials for large-scale in vitro screens.…”
Tunable biomaterials that mimic selected features of the extracellular matrix (ECM), such as its stiffness, protein composition, and dimensionality, are increasingly popular for studying how cells sense and respond to ECM cues. In the field, there exists a significant trade-off for how complex and how well these biomaterials represent the in vivo microenvironment, versus how easy they are to make and how adaptable they are to automated fabrication techniques. To address this need to integrate more complex biomaterials design with high-throughput screening approaches, we present several methods to fabricate synthetic biomaterials in 96-well plates and demonstrate that they can be adapted to semiautomated liquid handling robotics. These platforms include 1) glass bottom plates with covalently attached ECM proteins, and 2) hydrogels with tunable stiffness and protein composition with either cells seeded on the surface, or 3) laden within the three-dimensional hydrogel matrix. This study includes proof-of-concept results demonstrating control over breast cancer cell line phenotypes via these ECM cues in a semi-automated fashion. We foresee the use of these methods as a mechanism to bridge the gap between high-throughput cell-matrix screening and engineered ECM-mimicking biomaterials.
“…[48][49] The Lutolf group has demonstrated the use of 3D hydrogels in a microarray made using automated liquid handling robotics to test many microenvironmental conditions on the regulation of mouse embryonic stem cell self-renewal. 50 This study was a valuable demonstration of the utility of biomaterials in a high-throughput format and it was an important contribution towards using synthetic materials for large-scale in vitro screens.…”
Tunable biomaterials that mimic selected features of the extracellular matrix (ECM), such as its stiffness, protein composition, and dimensionality, are increasingly popular for studying how cells sense and respond to ECM cues. In the field, there exists a significant trade-off for how complex and how well these biomaterials represent the in vivo microenvironment, versus how easy they are to make and how adaptable they are to automated fabrication techniques. To address this need to integrate more complex biomaterials design with high-throughput screening approaches, we present several methods to fabricate synthetic biomaterials in 96-well plates and demonstrate that they can be adapted to semiautomated liquid handling robotics. These platforms include 1) glass bottom plates with covalently attached ECM proteins, and 2) hydrogels with tunable stiffness and protein composition with either cells seeded on the surface, or 3) laden within the three-dimensional hydrogel matrix. This study includes proof-of-concept results demonstrating control over breast cancer cell line phenotypes via these ECM cues in a semi-automated fashion. We foresee the use of these methods as a mechanism to bridge the gap between high-throughput cell-matrix screening and engineered ECM-mimicking biomaterials.
“…Traditional monolayer cultures of immortalized cancer cells lack original tumor heterogeneity (5,7,9), native histologic architectures, and cell-extracellular matrix interactions (22). Animal models of human cancers, including genetically engineered models and xenograft models, enable in vivo studies of cancer development, progression, and drug response.…”
Section: Primary Human Organoids As a Model Of Cancermentioning
Primary tumor organoids are a robust model of individual human cancers and present a unique platform for patient-specific drug testing. Optical imaging is uniquely suited to assess organoid function and behavior because of its subcellular resolution, penetration depth through the entire organoid, and functional endpoints. Specifically, optical metabolic imaging (OMI) is highly sensitive to drug response in organoids, and OMI in tumor organoids correlates with primary tumor drug response. Therefore, an OMI organoid drug screen could enable accurate testing of drug response for individualized cancer treatment. The objective of this perspective is to introduce OMI and tumor organoids to a general audience in order to foster the adoption of these techniques in diverse clinical and laboratory settings.
“…Even though these efforts have provided the research community with valuable insights into the mechanisms underlying a variety of biologic processes, it is nowadays widely accepted that knowledge obtained from these studies might be too reductionist to accurately translate to the human situation. [1,2] Growing cells onto 2D substrates deviates significantly from the dynamic three-dimensional (3D) in vivo situation; cells lack tissue-specific polarity, have limited contact with neighboring cells, and are exposed to non-physiologically uniform diffusion kinetics, which together alter how cells perceive and respond to their surrounding microenvironment (Fig. 1).…”
To gain a better understanding of the underlying mechanisms of neurological disease, relevant tissue models are imperative. Over the years, this realization has fuelled the development of novel tools and platforms, which aim at capturing in vivo complexity. One example is the field of biofabrication, which focuses on fabrication of three-dimensional (3D) biologically functional products in a controlled and automated manner. Herein, we provide a general overview of classical 3D cell culture platforms, particularly in the context of neurodegenerative disease. Subsequently, the focus is put on bioprinting-based biofabrication, its potential to advance 3D neuronal cell culture and, to conclude, the relevant translational bottlenecks, which will need to be considered as the field evolves.
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