NASH-in-a-box

Please click on the Request a quote link on this page and fill out the short form. A member of our expert team will contact you directly.

We do not provide healthy/lean media in the NASH-in-a-box kit as stellate cells cultured on plasticware become activated and generate a level of fibrosis regardless of the media. CN Bio’s NASH media, however, pushes the fibrosis response beyond baseline Stellate activation. This means that data normalized to a healthy/lean media control would be skewed versus a truly healthy liver microtissue. To circumvent this, CN Bio uses positive controls e.g., Elafibranor to compare therapeutic efficacy versus the negative control of the NASH phenotype.

To ensure that we recreate an in vitro model that is as human relevant as possible, cell lines (also known as immortalized cells), are not used. Instead, we use primary cells to generate physiologically relevant liver microtissues that more accurately recapitulate the disease phenotype.

Our cells are sourced from LifeNet Health LifeSciences, or Lonza and then thoroughly validated by CN Bio prior to inclusion in the kit. The process of validating cell lots for use in MPS assays is necessary to ensure data robustness and reliability. However, this process is costly and time-consuming. By providing customers with convenient, off-the-shelf, pre-validated reagents, we remove this burden – enabling researchers to focus their efforts on making new discoveries. Click here for more information about our wider portfolio of 3D validated cells and why cell validation ahead of organ-on-a-chip assays is critical to their success.

The model has been validated for 14 days of culture. The volumes of media and supplements supplied within the kit reflect this timeframe. Extending the experiment may be possible but has yet to be validated. Should you wish to register your interest, contact us.

Yes. We assess each qualified donor for key NASH-related genes. Please contact us to discuss your requirements.

NASH-in-a-box enables deep insights to be gathered from each microtissue, providing researchers with an in vitro experimental tool to study the mechanism of disease or anti-NASH drug treatments.

Approximately 1 mL of media can be removed from every media exchange. We recommend using the assay kits supplied to measure hepatic function and microtissue health (albumin & LDH) at each media exchange. Exchanged media can also be used to analyze soluble biomarkers of your choice throughout the experiment, such as interleukin-6 (IL-6) or tissue inhibitor of metalloproteinase I (TIMP-1). IL-6 is an excellent marker of the inflammatory response of NASH, while markers such as TIMP-1 and fibronectin can be used to assess fibrosis.

At the end of the experiment, microtissues can be recovered for microscopy analysis, quantitative PCR, and transcriptomics studies. The scaffold on which cells are seeded can be cut in half. One half of the recovered microtissue can be used for -omics or adenosine triphosphate (ATP) content quantification, and the other half can be fixed for immunofluorescence assays (IFA) using confocal microscopy.

A list of routinely used endpoint measurements can be found on our NASH applications page, however, we have tested many other disease biomarkers, so please contact us if your endpoint(s) of interest is/are not listed.

In an ideal world this would be the case, however, it is currently not possible to source all three cell types from the same donor. We thoroughly validate the performance of cells in tri-culture prior to supplying them in the kit.

You can find a sample validation report on this page that details the qualification protocol that we adhere to.

In short, primary human hepatocytes (PHHs) are first validated alone to verify 3D microtissue formation and acceptable function. Validated, PHHs are subsequently tested with Kupffer cells and stellate cells under NASH conditions to ensure good cell-cell interaction and disease phenotype using our bespoke NASH media.

We can only guarantee the performance of the NASH-in-a-box kit using the cells supplied. The cells within each kit have been thoroughly tested to ensure that they form 3D, highly functional microtissues that exhibit a NASH disease phenotype when cultured using our bespoke NASH media.

By using the kit, you benefit from years of CN Bio R&D expertise, validation of the 3D NASH model via industry collaboration, and carefully quality-controlled reagents that ensure reliable, robust, and reproducible performance. Our NASH-in-a-Box kit enables those who are new to the organ-on-a-chip (OOC) field to “hit the ground running” and fast-track their research.

However, should you want to develop your own 3D NASH model, or customize our tri-culture model to incorporate additional cell types, our PhysioMimix^® OOC range of microphysiological systems and Multi-chip Liver plates offer the flexibility and versatility to do so. Where possible, we would recommend purchasing cells from our 3D validated cell portfolio to shorten R&D timelines and minimize cost. Note: Stellate cells are only available for purchase within a NASH-in-a-box kit.

Our software guided protocol can be found on the PhysioMimix Controller Unit. It complements the Standard Operating Procedure (SOP) provided within the kit by providing step by step instructions for culturing over 14 days. The software will only allow the protocol to proceed once a user has confirmed that an action has been performed to prevent user error. By guiding users through the process, it enables any operator, irrespective of experience, to use the NASH-in-a-box kit with confidence.

NASH-in-a-box has been designed for, and validated using, the PhysioMimix OOC range of microphysiological systems. The Multi-chip Liver-12+ plates are for sole use with the NASH-in-a-box kit and are incompatible with other Organ-on-a-chip systems. Therefore, it would not make sense to purchase the kit for use with an alternative solution.

The PHHs supplied in the kit are from single donors that have been thoroughly validated for use in 3D organ-on-a-chip (OOC) cultures. Not all PHH donors cope well with 3D OOC cell culture or the NASH media. The former may explain why pooled hepatocytes generally perform poorly in organ-on-a-chip systems and is the reason why we provide 3D validated cells within our solutions portfolio.

As using pooled donors increases the risk of experimental failure, we recommend requesting different donor lots and testing these individually to overcome/assess donor to donor variability. The number of individual donors’ lots that we have in our validated tri-culture catalog varies depending on availability, however, we will do our best to accommodate your needs.

Our bespoke NASH media represents a fatty diet. It contains various free fatty acids (FFA) and sugars. Perfusion of liver microtissues with NASH media induces fat loading (steatosis) that leads to inflammation and fibrosis, thereby recapitulating human disease progression in vitro.

The recipe of media constituents is complex and has been developed over many years of R&D. When used in conjunction with PhysioMimix OOC Systems, NASH-in-a-box provides an off-the shelf solution to quickly, simply and confidently recreate the human NASH disease so that researchers can focus their time developing much-needed therapeutics for this widespread disease.

Shipping costs are not included as they may vary depending on the number of kits ordered or split shipment requests. Shipping costs will be added as a line item in any quote requests.

Included (separate shipment)

Not included