3D Cell Culture Bioreactor CERO 3D
The innovative 3D Cell Culture Incubator & Bioreactor

Stem cells – Spheroids – Organoids – Tissues

Highlights - 3D Cell Culture Incubator and Bioreactor CERO 3D

3D Cell Culture Bioreactor & Incubator CERO 3D
3D Cell Culture Bioreactor & Incubator CERO 3D
  • Improved viability and maturation
  • No embedding substrate required
  • Significantly reduced apoptosis & necrosis
  • No shear forces
  • Long-term cultivation for > 1 year
  • Significant reduced running cost

The CERO 3D Incubator & Bioreactor offers a unique 3D cell culture technology to boost your stem cell, spheroid, organoid and tissue research.

Highest levels of homogeneity and viability in long term cultures are just two benefits CERO 3D will provide.

Break the limits in 3D Cell Culture

The CERO 3D Incubator & Bioreactor is a new, revolutionary instrument creating optimal cell culture environment by monitoring and controlling temperature, pH and CO2 levels. 1- 4 individually controlled CEROtubes with a volume of up to 50ml provide highest biomass yields in a standardized and reproducible way, with minimum handling requirements. The CEROtubes, with small fins and a flat bottom, enable gentle cultivation conditions with reduced shear stress to your cells.

Pluripotent Stem Cells

The CERO 3D Incubator & Bioreactor provides the solution for scale-up and automation platforms, simplification and cost reduction of stem cell expansion projects in biobanks, cell-based drug discovery, toxicity testing and regenerative medicine.

  • Microcarrier-free
  • Stable pluripotency over many passages
  • Easy to set-up and simple workflow
  • Free-floating 3D aggregates
  • Able to differentiate in 3 germ layers
  • Homogeneous iPSC and ESC aggregates

Cero 3d Cell Culture Bioreactor iPSC
Human iPSC after expansion in CERO 3D

Pluripotent stem cells are directly inoculated as single cells into the CEROtube. During the self-aggregation the cells form homogeneous aggregates which can be expanded over many passages. The biomass increases significantly while only ~2min per day hands on time is required.

The resulting stem cell 3D aggregates can be processed directly for differentiation e.g. organoids or any other downstream application in 3D or 2D assay development.

CERO 3D Cell Culture Bioreactor iPSC Pluripotency
Human iPSC after expansion in CERO 3D Incubator & Bioreactor tested for pluripotency.
CERO 3D Cell Culture Bioreactor iPSC Differentiation
Human iPSC after expansion in CERO 3D Incubator & Bioreactor tested for differentiation in three germ layers.

Pluripotent stem cells expanded in CERO 3D (former name “BioLevitator”) will maintain pluripotency and can be differentiated into all 3 germ layers, as described by Elanzev et. al. 2015; Biotechnol. J. 2015, 10, 1589–1599:

Hepatitis Virus Research Model - Spheroids

The advent of long-term three dimensional cell culture holds a great promise in disease modeling and drug discovery. The cells kept in a 3D environment have the ability to mimic tissue-like structures more efficiently than in traditional 2D monolayer cultures. However, many scientist are struggling with many technical limitations when working with spheroids in long-term cultures.

The CERO 3D Incubator & Bioreactor is a revolutionary technology enabling scientists to perform experiments they were not able to do before.

  • No necrosis and apoptosis
  • Viability in long-term culture > 80 days
  • Improved maturation
  • Long-term proliferation
  • High homogeneity
  • High yield

Spheroids from HepG2 cells (hepatocyte cell line) cultivated in CERO 3D for >80 days.

Cells Proliferation
Cells are positive for cell proliferation marker KI67
Positive Cells
Cells are negative for apoptosis marker Casp.cl.3
Cells Albumin
Cells are positive for albumin

Virus Research

CERO 3D Incubator & Bioreactor enables growth and maturation of spheroids without necrosis and apoptosis while other technologies fail. Therefore, CERO 3D allows to maintain state-of-the-art 3D cultures of Huh7 hepatic cell line (see figure below). The disposable CEROtubes with Hepa filter allow safe virus experiments.

Virus Research CERO
Spheroid from hepatocyte cell line was matured for 20 days prior to exposure to HCV. Infection spreading (rate) was controlled after 24, 48 and 72hours (brown staining)

Cultivating hepatocyte spheroids in CERO considerably improves expansion, differentiation, maturation and hepatic virus infection compared to monolayer culture. Our research takes advantage from healthy cells even from long-term cultures in CERO. Moreover, we are now able to perform 3D long-term culture of human tissue specimen in CERO - a paradigm shift.

Prof. Dr. Heikenwälder, Chronic Inflammation and Cancer, Deutsches Krebsforschungszentrum (DKFZ), Heidelberg, Germany

Cardiac Tissue Model

Beating Cardiac Bodies – a complete workflow

Stem cell derived cardiomyocytes gain more and more attention in the field of cardiovascular research, disease modeling and drug development.

The CERO 3D Incubator & Bioreactor platform allows the workflow with stem cell expansion in homogeneous aggregates followed by direct induction into a high number of beating cardiac bodies. The expansion of pluripotent stem cells and subsequent cardiac induction/differentiation in CERO 3D results in a much higher cell quality, homogeneity, integrity and yield compared to traditional orbital shakers.

Cardiac Tissue Model
CERO 3D versus Orbital shaker – differentiation of murine embryonic stem cell derived cardiomyocytes 3, 8 and 13 days after cardiac induction.
  • Homogeneous 3D aggregates in suspension
  • Expansion and differentiation in the same CEROtube
  • Easy to handle & standardize
  • Highest yield
  • No substrate required
  • More relevance for drug treatments
  • More relevance for disease modeling

Purified and homogeneous cardiac bodies generated in CERO 3D Incubator & Bioreactor are a perfect tool for disease modeling and drug discovery. The pure cardiomyocytes can also be used in 2D downstream monolayer applications.

Moreover this 3D cultivation system substantially improves viability, maturation and contractility, therefore provides a reliable tool for cardiovascular therapies.

Yvonne Eibach, group of Prof. Thomas Braun, Max Planck Institute, Bad Nauheim

Organoids as Tumor Tissue Model

From Adult Stem Cells to Organoids

The CERO 3D Incubator & Bioreactor is a revolutionary technology for production of organoids from pluripotent or adult stem cells. It offers an efficient, standardizable way to generate and maintain high yields of homogeneous organoids used as a tool in cancer research. Remarkably, the organoids have also a predictive capacity for in vivo response.

  • Reduced costs and time
  • Increase size & lifespan
  • Optimal nutrition & gas diffusion
  • Free floating, no shear forces
  • High homogeneity and yield
  • Multiplexing organoidogenesis

Increase cellular, structural and functional complexity of organoids.

Gastric Organoids
Gastric organoids (bright field) at day 7 and after splitting on day 22 expanding from small cysts that expand to bigger spheres
Staining of Gastric Organoids
HE staining of Gastric organoids (2a) showing single layer of epithelia cells composed by the different cell types found in the stomach as seen in the PCR results (2b): expression of gastric mucins MUC5AC and MUC6, trefoil factors and pepsinogen, for instance. There are also stem cells markers like Lgr5 and Sox2.

The generation of gastric organoids is a crucial step in the study of Helicobacter pylori infection and gastric carcinogenesis. The CERO 3D Incubator & Bioreactor allows the generation in a much more efficient, reproducible, physiological and cost efficient way compared to any other approach

Dr. Raquel Mejias-Luque, TUM, Germany

Personalised and precision medicine

The progress in the development of molecular target therapies has improved the outcome in “in vivo”, however it is because of the tumor heterogeneity in patients that limits the efficiency of those drugs. Thus, to increase the relevance of “in vitro” models, it is particularly important to develop a reliable platform for growing primary tumor cells in a 3D in vitro model, like patient-derived xenografts or organoids. The CERO 3D Incubator & Bioreactor evolving a state-of-the-art dynamic culture system, accelerates precision medicine of primary tumor cells. The CERO 3D offers an efficient and reproducible approach to grow and maintain viable cancer cells in a CERO 3D Incubator & Bioreactor.

Positive Cholangiocellular
CK 20 positive Cholangiocellular carcinoma cancer cells
Viable Cholangiocellular
H & E staining: Viable Cholangiocellular carcinoma cancer cells
Positive Cells
MAPAC 155 positive cells

Our group is focusing on primary tumor cells grown on 3D polymer scaffolds. For that we are using CERO 3D to develop a model of drug development and precision medicine. It is particularly important that the cells are viable and proliferating in 3D complex structures as seen in tumor tissues. With the CERO 3D we are implementing and developing tools for patient-specific tumor therapy. Furthermore, the cancer cells isolated from fresh tumor biopsies make treatment optimization possible before a therapy is initiated. Here, CERO 3D incubator & bioreactor enables us to develop personalized medicine in a 3D cell culture model.

Prof. Dr. med. Felix Rückert, Geschäftsführender Oberarzt Chirurgische Klinik, Universitätsmedizin Mannheim

Meet the CERO

Upcoming CERO events

October
27th

EMBO Workshop: Microglia 2021

EMBO Workshop: Microglia 2021

Exhibition | EMBL Heidelberg, Meyerhofstraße 1, 69117 Heidelberg | 10 am CEST

Microglia are a resident macrophage of the central nervous system at the intersection of immunology and neuroscience. They have gained increasingly more attention over the past decade as more research shows that they are key contributors to neural circuit development and function. There is also increasing evidence that microglia are also important in the etiology of many nervous system diseases. This workshop will highlight the many important functions of microglia to nervous system physiology and function, the newest and most exciting literature and will bring together researchers using state-of the art imaging, genomics and molecular biology to study these enigmatic cells.


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