Ethical, scientific, and financial arguments make cell-based tests and three-dimensional cell cultures important tools to replace animal experiments. Cell culture devices have been developed that allow an in vitro environment with maximum retention of natural functionality. For this purpose, Greiner Bio-One has developed a 96 well high-throughput version of the well-established ThinCert® portfolio.
The new ThinCert® 96 well HTS insert is the optimal tool for scientists who want to generate tissue models (e.g., endothelia and epithelia) for transport, uptake, or co-culture studies in submersed as well as air lift culture with an automation-friendly format. The two-component system comprises a 96 well porous membrane plate manufactured entirely from polycarbonate (PC) and a corresponding polystyrene (PS) receiver plate. The tissue culture treated PC membrane allows for an ideal exchange of nutrients and substances, creating in vivo-like cultivation conditions to facilitate optimal cell growth, monolayer formation, and tissue differentiation.
The first variant of the new ThinCert® 96 well HTS insert product family includes two versions with a pore size of 0.4 µm. Further variants (3 µm, 8 µm) for migration and invasion assays will follow in 2023.
High-throughput in-vitro airway modelling with ThinCert® 96 Well HTS Insert - Application Note
Combining the Best of Two Worlds - ThinCert® HTS Insert whith Optimized Transparency (Item No. 655641)
By combining high membrane permeability and best possible transparency, Greiner Bio-One is offering a unique 96 well insert version, exclusively designed for applications that require both high diffusion rates as well as microscopic readouts. This is assured by a special pore arrangement and a high pore density of 2 x 107/cm2.
Proven Even Better - ThinCert® HTS Insert with Translucent Membrane (Item No. 655640)
In addition, a product version featuring a translucent membrane and a pore density of 1 x 108/cm2 complements the portfolio for experiments that need highest diffusion rates while imaging is not required.
What to look for in automation-friendly permeable cell culture supports
When choosing automation-friendly, permeable cell culture supports designed for high-throughput assays, look for these features:
Reduced wicking effect
A self-centering design results in a precisely centralized fit of the membrane plate that prevents capillary action (wicking) between the wall of the receiver well and the membrane insert for stable and reproducible assay conditions.
High transparency for microscopic inspections
When performing an assay that requires microscopic observation, it is essential to have a high level of transparency. The ThinCert® 96 well HTS insert membrane achieves this to a greater extent than its competitors while also maintaining a sufficient pore density for achieving the required permeability.
Enlarged basolateral access ports
This feature allows convenient access for (multichannel) pipettes, automated liquid handling robots as well as for electrodes of transepithelial electric resistance (TEER) measurement equipment. ThinCert® 96 well HTS inserts offer a 40% increased access area for more convenient liquid handling and TEER measurements compared to commercially available competitor products.
Superior membrane flatness
Superior flatness of the membranes guarantees optimal and reproducible cell culture conditions.
High-throughput in-vitro airway modelling with ThinCert® 96 Well HTS Insert - Application Note
ThinCert® HTS inserts and COVID research
Air-liquid interface (ALI) cell cultures have attracted even more attention during COVID disease modeling and are frequently applied for SARS-CoV-2 related research. There is an urgent need for a better understanding of COVID-19 pathogenesis, particularly host factors that facilitate viral infection and replication. Single permeable membrane inserts such as ThinCert® have proven to be an essential tool for modeling airway epithelia in vitro[1, 2, 3, 4].
Some examples: SARS-CoV-2 enters cells via binding to ACE2, followed by priming by TMPRSS2. Lukassen et al. examined the expression levels of ACE2 and TMPRSS2 and their distribution among different cell types in lung tissue and in cells from subsegmental bronchial branches by single nuclear and single cell RNA sequencing, respectively[1]. The researchers were able to collect data that provide a rich resource for future studies of COVID-19 infection and pathogenesis.
Djidrovski et al. developed a protocol to generate airway epithelial basal-like cells from induced pluripotent stem cells, which simplifies the manufacture of cellular models of the human upper airways. Basal-like cells were cultured on permeable inserts to allow formation of a confluent monolayer and then exposed to an air-liquid interface to induce differentiation into a pseudostratified epithelial construct with a marked similarity to the upper airway epithelium in vivo. The constructs contain the component cell types required of an epithelial model system, produce mucus and functional cilia, and can support SARS-CoV-2 infection/replication and the secretion of cytokines in a manner similar to that of in vivo airways. This method offers an accessible and highly scalable protocol for the manufacture of upper airway models for applications in development of therapies for respiratory viral infections and the assessment of drug toxicity on the human lungs [2].
ThinCert® HTS inserts for high-throughput assays
When scientists were performing in vitro experiments on cell lines in the 1990s, the idea that cell-based experiments would one day replace animal experiments was still a long way off and considered implausible by many scientists. Now, however, in the 2020s, we can exploit this potential – and in a high throughput manner, too!
With the new ThinCert® 96 well HTS inserts, Greiner Bio-One is expanding its well-established ThinCert® portfolio with an automation-friendly permeable membrane insert which has been optimised for high throughput assays. In contrast to single inserts, the insert plate can be handled as a single unit and significantly reduces manual handling and improves throughput.
Do you want to learn more about the potential of ThinCert® HTS inserts and how to use it in your cell culture research and high-throughput screening activities? Download our ThinCert® Application Note here!
High-throughput in-vitro airway modelling with ThinCert® 96 Well HTS Insert - Application Note