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Life Science
Incubator Subchamber
CERO 3D Incubator & Bioreactor
The CERO 3D Incubator & Bioreactor is a new, revolutionary instrument creating optimal cell culture environment. It offers a special 3D cell culture technology that monitors and controls temperature, pH and carbon dioxide levels. Indeed, this is ideal for stem cells, spheroids, organoids and even tissues.
The CERO 3D Incubator & Bioreactor provides some distinct advantages. First and foremost, it improves viability and maturation with minimized apoptosis and necrosis. The CERO 3D Incubator & Bioreactor has no shear forces, no requirement for embedding substrate, thus providing maximum homogeneity and allowing long-term cultivation for more than one year. Indeed, this will significantly reduce your running costs.
The individually controlled provide highest biomass yields. With volumes up to 50 ml, the CEROtubes, have small fins and a flat bottom allowing mild cultivation conditions in a standardized and reproducible way, with minimum handling requirements.
The CERO 3D Incubator & Bioreactor allows for simplified scale-up and automation platforms, and cost reduction. This is due to its distinct advantages of easy to set-up and simple workflows and minimum hands-on time that can be down to less than 2 minutes per day.
The CERO 3D Incubator & Bioreactor is indeed ideal for a wide variety of applications, from stem cell expansion projects in biobanks, cell-based drug discovery, toxicity testing and regenerative medicine, it is your ideal partner. For stem cells, The CERO 3D Incubator & Bioreactor is able to differentiate in 3 germ layers , provide homogeneous iPSC and ESC 3D aggregates. Your cells can be easily processed directly for differentiation such as for organoids or spheroids as a downstream application in 3D or 2D assay development and avoid technical limitations of long-term culture with high efficiency and standardization.
The CERO 3D Incubator & Bioreactor indeed is an ever-evolving a state-of-the-art dynamic culture system, accelerates your processes reduces costs and hands-on time and allows multiplexing. It provides optimal nutrition, gas diffusion thus increasing size and lifespan of your cultures.
Highlights - 3D Cell Culture Incubator and Bioreactor 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.
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Microcarrier-free
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Stable pluripotency over many passages
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Easy to set-up and simple workflow
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Free-floating 3D aggregates
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Able to differentiate in 3 germ layers
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Homogeneous iPSC and ESC aggregates
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.
Human iPSC after expansion in CERO 3D Incubator & Bioreactor tested for pluripotency.
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 are positive for cell proliferation marker KI67
Cells are negative for apoptosis marker Casp.cl.3
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.
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)
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.
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
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 (bright field) at day 7 and after splitting on day 22 expanding from small cysts that expand to bigger spheres
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.
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.
CK 20 positive Cholangiocellular carcinoma cancer cells
H & E staining: Viable Cholangiocellular carcinoma cancer cells
MAPAC 155 positive cells
| Protocols | Kindly refer to the "Downloads" tab. |
Bradley Justice, Nadia A.Badr, Robin A.Felder (2009)
Drug Discovery Today 102-107
3D cell culture technologies have revolutionized our understanding of cellular behavior, both in culture and in vivo, but adoption by cell-based screening groups has been slow owing to problems of consistency, scale and cost. The evolving field of high content screening technologies will, however, require a rethinking of 3D cell culture adoption to ensure the next generation of cells provide relevant in vivo-like data. Three current technologies are presented in this review: membranes, sponges/gels and microcarriers. A short history of these technologies and unique research applications are discussed. Also, the technologies are evaluated for usefulness in modern automated cell-based screening equipment.
Susan Breslin, Lorraine O’Driscoll
Drug Discovery Today Volume 240-249
Cells, grown as monolayers (2D models), are routinely used as initial model systems for evaluating the effectiveness and safety of libraries of molecules with potential as therapeutic drugs. While this initial screening precedes preclinical animal studies before advancing to human clinical trials, cultured cells frequently determine the initial, yet crucial, ‘stop/go’ decisions on the progressing of the development of a drug. Growing cells as three-dimensional (3D) models more analogous to their existence in vivo, for example, akin to a tumour, and possibly co-cultured with other cells and cellular components that naturally occur in their microenvironment may be more clinically relevant. Here, in the context of anti-cancer drug screening, we review 2D and 3D culture approaches, consider the strengths and relevance of each method.
Simone Allazetta, Tanja C. Hausherr, and Matthias P. Lutolf
Biomacromolecules 2013, 14, 4, 1122–1131
Droplet microfluidic technology is applied for the high-throughput synthesis via Michael-type addition of reactive, micrometer-sized poly(ethylene glycol) (PEG) hydrogels (“microgels”) with precisely controlled dimension and physicochemical properties. A versatile chemical scheme is used to modify the reactive PEG microgels with tethered biomolecules to tune their bioactive properties for the bioreactor culture and manipulation of various (stem) cell types.
Maria Mrakovcic, Markus Absenger, Regina Riedl, Claudia Smole, Eva Roblegg, Leopold F Fröhlich, Eleonore Fröhlich
PLoS One. 2013; 8(2);e56791
Nano-sized materials could find multiple applications in medical diagnosis and therapy. One main concern is that engineered nanoparticles, similar to combustion-derived nanoparticles, may cause adverse effects on human health by accumulation of entire particles or their degradation products. Chronic cytotoxicity must therefore be evaluated. In order to perform chronic cytotoxicity testing of plain polystyrene nanoparticles on the endothelial cell line EAhy 926, we established a microcarrier cell culture system for anchorage-dependent cells (BioLevitatorTM). Cells were cultured for four weeks and exposed to doses, which were not cytotoxic upon 24 hours of exposure. For comparison, these particles were also studied in regularly sub-cultured cells, a method that has traditionally been used to assess chronic cellular effects. Culturing on basal membrane coated microcarriers produced very high cell densities. Fluorescent particles were mainly localized in the lysosomes of the exposed cells. After four weeks of exposure, the number of cells exposed to 20 nm polystyrene particles decreased by 60% as compared to untreated controls. When tested in sub-cultured cells, the same particles decreased cell numbers to 80% of the untreated controls. Dose-dependent decreases in cell numbers were also noted after exposure of microcarrier cultured cells to 50 nm short multi-walled carbon nanotubes. Our findings support that necrosis, but not apoptosis, contributed to cell death of the exposed cells in the microcarrier culture system. In conclusion, the established microcarrier model appears to be more sensitive for the identification of cellular effects upon prolonged and repeated exposure to nanoparticles than traditional sub-culturing.
Zhen-Yu Yang, Jun-Tao Kan, Ze-Yu Cheng, Xian-Li Wang, Yi-Zhun Zhu & Wei Guo
Cytotechnology volume 66, pages51–61(2014)
Daphnoretin is a bicoumarin compound isolated from a natural product, Wikstroemia indica, which has been used to treat many diseases. It has strong antiviral and anti-tumor activities. Taking the anti-tumor activity of daphnoretin as a starting point, the present study aimed to test the pro-apoptotic effect of daphnoretin and its underlying mechanism in HeLa cells. The inhibitory effects of daphnoretin on viability and proliferation of HeLa cells were determined by the MTT assay. Daphnoretin-induced apoptotic morphological changes were analyzed by mitochondrial membrane potential and Hoechst staining. The number and stage of apoptotic HeLa cells were determined by flow cytometry. Gene expression was determined by reverse-transcription polymerase chain reaction. Protein expression was determined by western blot. The caspase activity of HeLa cells was detected by a caspase-3 and caspase-9 colorimetric assay kit. We found that daphnoretin significantly inhibited HeLa cells’ viability by the MTT assay and flow cytometry. The nuclei of the apoptotic cells exhibited strong, blue fluorescence in Hoechst staining. Bax mRNA and protein levels were increased while bcl-2 mRNA levels were decreased after daphnoretin treatment. Daphnoretin also activated both caspase-3 and caspase-9. These findings suggest that daphnoretin promotes apoptosis of HeLa cells in a mitochondria-mediated way. Daphnoretin therefore has potential to be a promising drug to treat uterine cervix cancer.