3D Cell Culture Technology 

CelVivo ClinoStar System - Mimicking in vivo performance

The CelVivo ClinoStar System creates a cell culture environment which promotes the growth, maintenance and functionality of large 3D tissue mimetic structures, whether they are spheroids, organoids or other aggregates. 

In this environment, liver spheroids for example remain in a stable dynamic equilibrium and exhibit human in vivo physiological performance for at least 24 days (i.e. up to at least 42 days of culture). During this time, the spheroids exhibit a stable transcriptome profile. 

• Large spheroids created with high uniformity in size
• Excellent for co-cultures and tumor studies
• Cells experience very low shear stress
• Active diffusion enables improved nutrient and gas exchange
• Function, architecture and ultrastructure of 3 week old spheroids mimic those seen in vivo

ClinoStar Advanced Bioreactor Platform

The ClinoStar System employs a clinostat principle (a rotating bioreactor) to keep the cells in suspension by counterbalancing the gravitational forces. This approach has the advantage that the spheroids and organoids formed are exposed to very low shear forces and benefit from improved gas exchange and nutrient supply. Low shear forces, combined with active diffusion, are key parameters for cells to develop into functional 3D constructs. Two components build the ClinoStar System, the ClinoStar Incubator and the ClinoReactor.

ClinoStar Incubator for 3D Cell Culture

The ClinoStar Device is a clinostat CO₂ incubator which allows you to culture cells in 3D. The easily adjustable rotation of the bioreactor, see below, not only abolishes the force of gravity on the cells but also leads to a much better gas exchange and nutrition supply of the cells. A tablet with an intuitive software is used to control the temperature, CO₂ level, and each of the six clinostat motors (independently). A centrally located fan blends the CO₂ and temperature in the chamber to ensure a precise, homogeneous growth environment. 

ClinoReactor All-in-One Bioreactor for ClinoStar

The ClinoReactor is an all-in-one bioreactor. The front wall of the chamber is also a lid which can be clicked on and off as desired giving full access to the chamber in a petri-dish manner. The vents in top of the ClinoReactor ensure a constant flow of air over the membrane allowing CO₂ to be in equilibrium with the media inside the cell chamber. In addition, the reactors have their own humidification system for minimizing the risk of infections and for keeping the volume constant in the incubation chamber.

How It Works

Culturing spheroids and organoids is dependent on the cell model and sample being cultured. The ClinoStar System is a generic system to generate spheroids and organoids in a reproducible fashion. The initial cell aggregation can be performed in several ways to accommodate various cell and sample types, the downstream process remains.

Spheroid and Organoid Development 

1. Select aggregation method: Following 2D cell expansion or tissue disaggregation, the cells can be seeded in the ClinoReactor with one of three approaches: as a single cell suspension, or following initial aggregation preformed using a hydrogel or in micropattern plates.

2. Transfer to ClinoReactor: When starting with the ClinoStar system, you can use the cell culture media and supplements that you are used to. Specialized media, scaffolds, ECM substitutes or growth factors are not needed. 

3. Cultivate in ClinoStar: In the cultivation phase, cell culture media is routinely renewed, and irregular constructs are removed from the ClinoReactor to ensure uniform growth. Check that your constructs are functional against an in vivo benchmark that can be assayed in vitro.

4. Mature spheroids or organoids: When the cell lines have recovered their functionality, they are considered mature and ready for experimentation. This occurs after 18 days for HEPG2/C3A cells. At that time, each ClinoReactor will hold around 350 spheroids, each containing about 82,000 cells.