4 research outputs found
Lamination And Microstructuring Technology for a Bio-Cell Multiwell array
Microtechnology becomes a versatile tool for biological and biomedical
applications. Microwells have been established long but remained
non-intelligent up to now. Merging new fabrication techniques and handling
concepts with microelectronics enables to realize intelligent microwells
suitable for future improved cancer treatment. The described technology depicts
the basis for the fabrication of a elecronically enhanced microwell. Thin
aluminium sheets are structured by laser micro machining and laminated
successively to obtain registration tolerances of the respective layers of
5..10\^Am. The microwells lasermachined into the laminate are with
50..80\^Am diameter, allowing to hold individual cells within the well.
The individual process steps are described and results on the microstructuring
are given.Comment: Submitted on behalf of EDA Publishing Association
(http://irevues.inist.fr/EDA-Publishing
MatriGridÂź based biological morphologies: tools for 3D cell culturing
Recent trends in 3D cell culturing has placed organotypic tissue models at another level. Now, not only is the microenvironment at the cynosure of this research, but rather, microscopic geometrical parameters are also decisive for mimicking a tissue model. Over the years, technologies such as micromachining, 3D printing, and hydrogels are making the foundation of this field. However, mimicking the topography of a particular tissue-relevant substrate can be achieved relatively simply with so-called template or morphology transfer techniques. Over the last 15 years, in one such research venture, we have been investigating a micro thermoforming technique as a facile tool for generating bioinspired topographies. We call them MatriGrid Âź s. In this research account, we summarize our learning outcome from this technique in terms of the influence of 3D micro morphologies on different cell cultures that we have tested in our laboratory. An integral part of this research is the evolution of unavoidable aspects such as possible label-free sensing and fluidic automatization. The development in the research field is also documented in this account
Organ-on-a-Disc: A Scalable Platform Technology for the Generation and Cultivation of Microphysiological Tissues
Organ-on-Chip (OoC) systems culture human tissues in a controllable environment under microfluidic perfusion and enable a precise recapitulation of human physiology. Although recent studies demonstrate the potential of OoCs as alternative to traditional cell assays and animal models in drug development as well as personalized medicine, unmet challenges in device fabrication, parallelization and operation hinder their widespread application. In order to overcome these obstacles, this thesis focuses on the development of the Organ-on-a-Disc technology for the scalable generation and cultivation of microphysiological tissues. Organ-Discs are fabricated using precise, rapid and scalable microfabrication techniques. They enable the pump- and tubing-free perfusion as well as the parallelized generation and culture of tailorable and functional microtissues using rotation-based operations. The Organ-Disc setup is suitable for versatile tissue readouts, treatments and even whole blood perfusion with minimal handling and equipment requirements. Overall, the Organ-Disc creates a scalable and userfriendly platform technology for microphysiological tissue models and paves the way for their transition towards high-throughput systems.:Abbreviations
Symbols
1 Introduction
2 Background
2.1 Fluid Dynamics
2.1.1 Flow Equations
2.1.2 Hydraulic Resistance
2.1.3 Wall Shear Stress
2.1.4 Centrifugal Microfluidics
2.2 Microfluidic Chip Fabrication
2.2.1 Chip Materials
2.2.2 Microstructuring
2.2.3 Bonding
3 State of the Art
3.1 Cell Culture Systems
3.2 3D Tissue Generation in Microfluidic Systems
3.3 Organ-on-Chip
3.4 Scale-up of Organ-on-Chip Systems
3.4.1 Scalable Fabrication Technologies
3.4.2 Parallelization Approaches
3.4.3 Integrated Fluid Actuation
3.5 Centrifugal Microfluidics
4 Objectives
5 Materials and Methods
5.1 Organ-Disc Fabrication
5.1.1 Materials
5.1.2 2D Structuring
5.1.3 Hot Embossing
Stamp Fabrication
TPE Hot Embossing
5.1.4 Bonding
Solvent Vapor Bonding
Thermal Fusion Bonding
TPE Bonding
5.1.5 Characterization Methods
Structure Sizes
Bonding Strength
Optical Properties
5.2 Organ-Disc Spinner
5.2.1 Centrifugal Loading Setup
5.2.2 Centrifugal Perfusion Setup
5.2.3 Peristaltic Pumping Setup
5.3 Organ-Disc Perfusion
5.3.1 Centrifugal Perfusion
5.3.2 Peristaltic Perfusion
5.4 Preparatory Cell Culture
5.5 Organ-Disc Cell Loading
5.5.1 Centrifugal Cell Loading
5.5.2 Endothelial-lining
5.6 Organ-Disc Cell Culture
5.6.1 Staining and Imaging
Live Cell Labeling
Live/Dead Staining
CD106 Staining
CD41 Staining
Fixation, Permeabilization and Blocking
Actin/Nuclei Staining
CD31/Nuclei Staining
5.6.2 Media Analysis
5.6.3 Endothelial Cell Activation
5.6.4 Whole Blood Perfusion
5.7 Data Presentation and Statistics
6 Concept and Design
6.1 Organ-Disc Technology
6.2 Organ-Disc Design
6.3 Centrifugal Cell Loading
6.4 Endothelial Cell Lining
6.5 Centrifugal Perfusion
6.6 Peristaltic Perfusion
7 Building Blocks
7.1 Microfabrication Technology
7.1.1 Structuring
2D Structuring
Hot Embossing
7.1.2 Bonding
Solvent Vapor Bonding
Thermal Fusion Bonding
TPE Bonding
7.2 Organ-Disc Spinner
8 Perfusion
8.1 Centrifugal Pumping
8.2 Peristaltic Pumping
9 Tissue Generation and Culture
9.1 3D Tissue Generation
9.2 Stratified Tissue Construction
9.3 Generation of Endothelial-lined Channels
9.4 Perfusion of Endothelial-lined Channels
9.4.1 Media Monitoring
Evaporation
Cell Metabolism
9.4.2 Inflammatory Cell Stimulation
9.4.3 Whole Blood Perfusion
10 Discussion
10.1 Organ-Disc Technology
10.2 Scalable, Precise and Robust Organ-Disc Fabrication
10.2.1 Fabrication of Thermoplastic Organ-Discs
10.2.2 Fabrication of TPE Modules
10.2.3 Integration of TPE Modules to Organ-Discs
10.3 Tunable, Pump- and Tubing-free Perfusion
10.4 On-Disc Tissue Culture
10.4.1 3D Tissues
10.4.2 Blood Vessel-like Structures
10.4.3 Tissue Characterization and Treatment
10.5 On-Disc Blood Perfusion
11 Summary and Conclusion
12 References
13 AppendixIn Organ-on-Chip (OoC)-Systemen werden menschliche Gewebe mittels mikrofluidischer Versorgung in einer kontrollierten Umgebung kultiviert und so die Physiologie des Menschen nachgebildet. Obwohl aktuelle Studien zeigen, dass dieser Ansatz Alternativen zu herkömmlichen Zellbasierten Tests und Tiermodellen in der Arzneimittelentwicklung und der personalisierten Medizin bietet, stehen einer breiteren Anwendung HĂŒrden im Bereich der Herstellung, Parallelisierung und Handhabung im Weg. Deshalb ist das Ziel dieser Arbeit die Entwicklung der Organ-on-a-Disc-Technologie, die eine skalierbare Erzeugung und Kultur von mikrophysiologischen Geweben ermöglicht. FĂŒr die Herstellung von der Organ-Disc kommen prĂ€zise, schnelle und skalierbare Mikrofabrikationsmethoden zum Einsatz. Die Organ-Disc schafft die Basis fĂŒr die parallelisierte Erzeugung und Kultur von maĂgeschneiderten und funktionellen Mikrogeweben, sowie deren Versorgung durch rotationsbasierte Prozesse und ohne zur Hilfenahme von Pumpen oder SchlĂ€uchen. Die Organ-Disc eignet sich fĂŒr unterschiedliche Charakterisierungsmethoden sowie der Gewebestimulation und sogar der
Vollblutperfusion mit minimalem Aufwand und Equipment. Insgesamt stellt die Organ-Disc eine skalierbare und benutzerfreundliche Plattformtechnologie fĂŒr mikrophysiologische Modelle dar und bereitet den Weg fĂŒr Hochdurchsatzanwendungen.:Abbreviations
Symbols
1 Introduction
2 Background
2.1 Fluid Dynamics
2.1.1 Flow Equations
2.1.2 Hydraulic Resistance
2.1.3 Wall Shear Stress
2.1.4 Centrifugal Microfluidics
2.2 Microfluidic Chip Fabrication
2.2.1 Chip Materials
2.2.2 Microstructuring
2.2.3 Bonding
3 State of the Art
3.1 Cell Culture Systems
3.2 3D Tissue Generation in Microfluidic Systems
3.3 Organ-on-Chip
3.4 Scale-up of Organ-on-Chip Systems
3.4.1 Scalable Fabrication Technologies
3.4.2 Parallelization Approaches
3.4.3 Integrated Fluid Actuation
3.5 Centrifugal Microfluidics
4 Objectives
5 Materials and Methods
5.1 Organ-Disc Fabrication
5.1.1 Materials
5.1.2 2D Structuring
5.1.3 Hot Embossing
Stamp Fabrication
TPE Hot Embossing
5.1.4 Bonding
Solvent Vapor Bonding
Thermal Fusion Bonding
TPE Bonding
5.1.5 Characterization Methods
Structure Sizes
Bonding Strength
Optical Properties
5.2 Organ-Disc Spinner
5.2.1 Centrifugal Loading Setup
5.2.2 Centrifugal Perfusion Setup
5.2.3 Peristaltic Pumping Setup
5.3 Organ-Disc Perfusion
5.3.1 Centrifugal Perfusion
5.3.2 Peristaltic Perfusion
5.4 Preparatory Cell Culture
5.5 Organ-Disc Cell Loading
5.5.1 Centrifugal Cell Loading
5.5.2 Endothelial-lining
5.6 Organ-Disc Cell Culture
5.6.1 Staining and Imaging
Live Cell Labeling
Live/Dead Staining
CD106 Staining
CD41 Staining
Fixation, Permeabilization and Blocking
Actin/Nuclei Staining
CD31/Nuclei Staining
5.6.2 Media Analysis
5.6.3 Endothelial Cell Activation
5.6.4 Whole Blood Perfusion
5.7 Data Presentation and Statistics
6 Concept and Design
6.1 Organ-Disc Technology
6.2 Organ-Disc Design
6.3 Centrifugal Cell Loading
6.4 Endothelial Cell Lining
6.5 Centrifugal Perfusion
6.6 Peristaltic Perfusion
7 Building Blocks
7.1 Microfabrication Technology
7.1.1 Structuring
2D Structuring
Hot Embossing
7.1.2 Bonding
Solvent Vapor Bonding
Thermal Fusion Bonding
TPE Bonding
7.2 Organ-Disc Spinner
8 Perfusion
8.1 Centrifugal Pumping
8.2 Peristaltic Pumping
9 Tissue Generation and Culture
9.1 3D Tissue Generation
9.2 Stratified Tissue Construction
9.3 Generation of Endothelial-lined Channels
9.4 Perfusion of Endothelial-lined Channels
9.4.1 Media Monitoring
Evaporation
Cell Metabolism
9.4.2 Inflammatory Cell Stimulation
9.4.3 Whole Blood Perfusion
10 Discussion
10.1 Organ-Disc Technology
10.2 Scalable, Precise and Robust Organ-Disc Fabrication
10.2.1 Fabrication of Thermoplastic Organ-Discs
10.2.2 Fabrication of TPE Modules
10.2.3 Integration of TPE Modules to Organ-Discs
10.3 Tunable, Pump- and Tubing-free Perfusion
10.4 On-Disc Tissue Culture
10.4.1 3D Tissues
10.4.2 Blood Vessel-like Structures
10.4.3 Tissue Characterization and Treatment
10.5 On-Disc Blood Perfusion
11 Summary and Conclusion
12 References
13 Appendi
Physical Aspects of Cell Culture Substrates: Topography, Roughness, and Elasticity
The cellular environment impacts a myriad of cellular functions by providing signals that can modulate cell phenotype and function. Physical cues such as topography, roughness, gradients, and elasticity are of particular importance. Thus, synthetic substrates can be potentially useful tools for exploring the influence of the aforementioned physical properties on cellular function. Many microâ and nanofabrication processes have been employed to control substrate characteristics in both 2D and 3D environments. This review highlights strategies for modulating the physical properties of surfaces, the influence of these changes on cell responses, and the promise and limitations of these surfaces in inâvitro settings. While both hard and soft materials are discussed, emphasis is placed on soft substrates. Moreover, methods for creating synthetic substrates for cell studies, substrate properties, and impact of substrate properties on cell behavior are the main focus of this review. The cellular environment plays a significant role in cell phenotype and function. As such, physical properties of cell culture substrates including topography, roughness, and elasticity may be utilized to investigate the influence of these physical cues on the cellular response. In this review, strategies for modulating the physical properties of surfaces, the influence of these changes on cell responses, and the promise and limitations of these surfaces in inâvitro settings are highlighted, with a particular emphasis on elastic substrates.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/90132/1/336_ftp.pd