Organ-on-a-Chip - LiveBox2
- Product SKU: IV-K02-EU-PET
- Category: Cell Culture Chips, Microfluidic Chips, Microfluidic Organ-on-a-chip
LB2 is an Organ-on-a-Chip platform characterized by a transparent bioreactor divided in two connected chambers to host 2D or 3D cell cultures and tissues (e.g. biopsies). The two chambers are designed to develop models of physiological barriers and to ensure an in vivo-like environment under static or dynamic flow conditions thanks to the presence of 2 flow inlets and 2 outlets for the perfusion of culture media and testing solutions.
The setup includes all you need to perform your experiment: a LB2 bioreactor with its Clamp, Porous Membranes, Tubes, Connectors and Mixing Chambers for media and reagents.
LiveBox2 is an innovative bioreactor developed by IVTech, an italian biotech company whose aim is to provide a new kind of in vitro models based on dynamic fluid flows. Darwin Microfluidics as partner of IVTech offers the same purchase conditions together with a scientific support in 3D cell culture and microfluidics.
This device consists in two inter-connected culture chambers and it allows a precise and continuous control of culturing parameters. Thanks to the presence of a removable holder, the bioreactor can host a porous membrane to model in vitro epithelial barriers such as lung and intestinal mucosa and endothelial barrier. It is possible to mimic tissue organization, cell-cell interactions and physiological responses to external stimuli by maintaining the sample in a dynamic environment.
We provide different porous membranes. Let us to guide you in choosing the best solution for your needs:
|Material||Pore sizes, densities and thicknesses||Cell culture treated||Optical property||Suggested applications|
|Polyethylene terephthalate (PET)||
||Yes||Transparent (translucent with 25 µm thickness)||
A standard microscope Glass Disc Coverslip (Φ 20 mm and thickness 0.17 mm) represents the bottom support of the device. It is possible to seed the cells on it as well. It can be 1) easily placed into the chamber and 2) removed from it at the end of your experiment. The Glass Disc Coverslip is optically transparent and make the bioreactor suitable for microscopic imaging.
Two integrated inlets and outlets together with the clamp system ensure leak-free junctions in both static and dynamic experiments (flow rate up to 0.5 mL/min in the apical or basal compartment).
All the components of the bioreactor are made in medical silicone PE500, so the chambers can be used several times (you have just to replace the glass coverslip and the porous membrane) and tolerates different washing and sterilization cycles in autoclave.
Features & Advantages
The Organ-on-a-Chip LB2 is designed to offer a reversible access to the cultured cells and tissues.
The bioreactor has been designed to reproduce the typical surface of a 12-well transwell to allow you to use your standard protocols, cell dilutions and common cell culture settings.
The bottom glass support makes the microscopic imaging convenient:
Real-time live imagining of the 2D/3D cell culture during the static or dynamic experiments by using bright-field, fluorescence or confocal microscopes.
Down-stream staining and imaging analysis; since the glass support is removable, you have the possibility to disassemble the chamber and remove the glass disc with the tissue! Use it for further investigations such as high-resolution imaging.
The removable holder is placed at the interconnection between the 2 chambers:
It is designed to host porous membranes with a diameter of 25 mm.
It allows to handle the membrane easily and safely even when cells are seeded.
Post-culture imaging and downstream analysis are possible. Once the LB2 is disassembled, the holder system supporting the cultivated cell monolayer or 3D construct can be easily removed and used for further investigations and analysis.
The interconnection of different bioreactors can create a modular system to mimic the cross-talk between different tissues in a Body-on-a-Chip configuration.
Components of the platform:
- LiveBox2 bioreactor equipped with glass discs
- Porous membranes
- Membrane holder
- Silicon tubes with Luer Lock connectors
- Mixing chambers / reservoirs with their support
- Extractor to easily remove the glass disc
The culture chamber, the glass discs and the clamp, once assembled together, create an optically transparent bioreactor with a perfect sealing up to flow rates of 0.4 mL/min. The porous membrane and its holder have to be placed at the interface between the two chambers. The 2 inlets and 2 outlets allow medium perfusion and dynamic flow conditions during your experiments of drug screening and/or cell differentiation by creating 2 independent fluidic circuits.
LB2 is compatible with commercial peristaltic pumps (click here to see our large offer!) and with the innovative LiveFlow Pump that can be placed directly in the wet environment of your cell incubator (click here for more information!).
1x LB2 bioreactor
|Material||Medical grade silicone PE500|
|Cell culture bottom support||8x Glass Disc Coverslip (Φ 20 mm and thickness 0.17 mm)|
|Chamber Volume||1.5 mL|
|2 Inlet/Outlet||4x Luer Locks M/F (polypropylene)|
|Suggested Flow Rate||0.1 to 0.4 mL/min|
1x Clamp System
|Material||Derlin and Acrylonitrile butadiene styrene (ABS)|
|Window size||Φ 20 mm|
2x Mixing Chambers
|Material||ABS; Stainless steel spine|
|Component||A support to safely remove the cell culture chamber from the clamp system at the end of the experiment; it is to avoid the break the Glass Disc|
|Material||Medical Silicone PE500|
1x Holder System
|Dimensions of the suggested membrane||
1x Pack of 100 Porous Membranes
|Material||To choose between: Polyethylene terephthalate (PET), Nylon or Polycarbonate|
|Pore size||0.4 µm - 100 µm|
The individual components are available upon request.
The complete setup comes sterile and have to be stored in dry conditions, without direct exposition to the sunlight at room temperature (15-25 °C).
Let us to guide you in choosing the best solution for your needs!
IVTech technology is based on compact, user-friendly and transparent cell culture chambers with shape and dimensions similar to the classical 24 well plate (1.5 mL/culture chamber), widely used in biological research. The holder can host membranes with a traditional size of a 12-well transwell. This allow you to use the same standard protocols of your habitual cell culture experiments with the advantages to create 3D tissues and a dynamic environment.
The design and dimensions of the LiveBox2 make it suitable for the direct creation of a 3D complex tissue through the use of a 3D bioprinter!
Seed your cell line on the standard glass disc and/or on the porous membrane. Once the assembly is completed, the two-chamber device is hermetically closed and ready to work in dynamic conditions. After clamping, solutions can be injected to recreate the physiological conditions or test the effects of drugs and growth factors. Many configurations can be set by using the 2 inlets and 2 outlets as show in the image below:
- Tangential flow configuration, suitable for physiological barriers simulation. The apical and basal chambers are perfused by 2 different and independent flows. The flow lines are parallel to the membrane surface.
- Perfusion flow configuration, suitable for 3D cells construct perfusion. Flow lines cross the cells construct in the vertical direction. Two sub-configurations can be implemented, based on the flow direction: top-down or down-top configurations.
Tissues inside the bioreactor can be observed by microscopy or harvested for downstream analysis.
- Reversible glass cosverslip integration thanks to the modular bioreactor
- Leak-free Luer Connections
- Highly-resistant and optically clear materials
- Dimensions compatibile with microscopes observation
Examples of application
This modular platform significantly improve the predictability of drug screening and studies of personalized medicine.
LiveBox2 has been successfully used to create the following tissues:
- Physiological models: Bone, Intestinal Epithelium, Kidney, Lung
- Disease models: Metastasis
- Multi-organ models: Gastro-Intestinal duct
Useful cell culture protocols can be find in these papers published in high impact factor journals:
Case study n° 1: Biopsy (muscle explant)
Figure 1. Soleus muscle, samples maintained in culture under fluid dynamic (a,c,e) or conventional (b,d,f ) conditions. After 24 h incubation in the bioreactor (c), cytoskeletal organization is still recognizable, and mitochondria (arrows), sarcoplasmic reticulum (arrowheads) and glycogen (thin arrows) are well preserved; under conventional condition (d) the cytoskeletal architecture is loosened, mitochondria (arrows) and sarcoplasmic reticulum (arrowheads) are swollen and glycogen is lost. After 48 h in the bioreactor (e), sarcomere organization is hardly recognizable, sarcoplasmic reticulum (arrowheads) is swollen, but mitochondria (arrows) are still preserved; under conventional condition (f ), the cytoskeletal organization is definitely lost, and mitochondria (arrows) and sarcoplasmic reticulum (arrowheads) are strongly damaged. Scale bars: 1000 nm. Insets scale bars: 10 μm.
Case study n° 2: Breast Cancer Model (drug cytotoxicity)
Figure 2. A 3D culture of MCF7 is more resistant than 2D culture to the treatment with Trastuzumab. However, both culture conditions show a higher sensitivity when the drug test is performed in a dynamic environment. The fluid flow provide a prediction closer to the in vivo drug efficacy than the static models.
Case study n° 3: Obesity Model (multi-organs)
Figure 3. a) Basic components of the 3-way fluidic system with LB1 and LB2 chambers; b) fluidic setup with peristaltic pump, mixing chamber and hepatic (LB1), endothelial and adipose tissue (LB2) chambers; c) HepG2 cells stained with DAPI (blue) and actin-phalloidin (red) seeded on porous collagen scaffolds (green autofluoresence). Scale bar 50 μm; d) endothelial cells stained with calcein. Scale bar 50 μm; e) adipocytes stained with oil-red. Scale bar 50 μm.
- Performing an experiment under dynamic flow conditions is possible in many different ways, it depends on the type of results you aim to obtain:
- If your experiment requires a short exposure time you can perform it without incubator by just keeping the medium at the right temperature and using CO2 independent medium (Leibovitz L-15 for example);
- If your experiment requires a prolonged incubation you can use the innovative LiveFlow Pump that can be placed directly in the wet environment of your cell incubator together with the bioreactor (click here for more information!).
- The sterilization of the components is possible trough an autoclave or by using a solution of 70% ethanol.
- It would be possible to reuse the chip after washing and sterilization. Only the Glass Discs and the membrane have to be changed after each utilization.
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