Organ-on-a-Chip - LiveBox2
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Organ-on-a-Chip - LiveBox2

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. 
Available within 2-3 weeks
€636.60
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. 

Description

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)
  • Pore size 0.45 µm, pores density 2*10^6, thickness 12 µm
  • Pore size 1 µm, pores density 2*10^6, thickness 11 µm
  • Pore size 3 µm, pores density 8*10^5, thickness 9 µm
  • Pore size 8 µm,  pore density 6*10^4, thickness 16 µm
Yes Transparent
  • Permeability studies
  • Cell culture in adhesion
  • Microscopic observation
Polycarbonate (PC)
  • Pore size 3 µm,  pores density 1.6*10^6, thickness 22 µm
  • Pore size 8 µm,  pores density 1*10^5, thickness 12 µm
  • Pore size 0.4 µm, pores density 1.5*10^8, thickness 25 µm
Yes Transparent (translucent with 25 µm thickness)
  • Permeability studies
  • Cell culture in adhesion
  • Microscopic observation (not possible for the 25 µm thickness)
Nylon
  • Square mesh, mesh layer 20 µm, porosity 14%, hydrophilic
  • Square mesh, mesh layer 30 µm, porosity 17%, hydrophilic
  • Square mesh, mesh layer 41 µm, porosity 31%, hydrophilic
  • Square mesh, mesh layer 100 µm, porosity 44%, hydrophilic
No Opaque (white)
  • Scaffold perfusion (scaffold placed in LB2 apilcal chamber)
  • Spheroids or cells aggregates in floating conditions (placed in LB2 bottom chamber)

 

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
  • Clamp
  • 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
Components
  • 2x Support Columns
  • 1x Base (ABS)
  • 1x Low Friction Ring
  • 1x S-shape Upper Component (ABS)
 

2x Mixing Chambers

Material Polypropylene; ABS
Components
  • 2x Medium reservoir 33 mL
  • 6x Luer Locks M/F
  • 2x On/Off valve
  • 2x Mixing Chamber Support
 

1x Extractor

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
 

8x Tubes

Material Medical Silicone PE500
Components
  • 8x Tubes of 20 cm each
  • 16x Luer Locks M/F (polypropylene)


1x Holder System

Material Derlin 
Dimensions of the suggested membrane
  • Φ 25 mm
  • Thickness up to 1 mm
Components
  • 1x Core
  • 1x Shield

1x Pack of 100 Porous Membranes

Material  To choose between: Polyethylene terephthalate (PET), Nylon or Polycarbonate 
Dimensions
  • Φ 25 mm
  • Thickness up to 1 mm
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!

Setup

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:

  1. 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.
  2. 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. 

 

Benefits:

  • 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)

    Carton et al., Incubation under fluid dynamic conditions markedly improves the structural preservation in vitro of explanted skeletal muscles. Eur J Histochem, 2017, 61(4): 2862 (PDF file)

      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)

      Civita P., Drug response assay on Transtuzumab sensitive breast cancer cell lines in a dynamic 3D culture system. Oral presentation: SIBBM 2017, Frontiers in molecular biology. June 2017 Milan 

       

      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)

      Ahluwalia A. et al., Systemic and vascular inflammation in an in-vitro model of central obesity. PlosOne, 2018, 13(2): e0192824 (PDF file)

       

      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.

      

      User guide

      • Performing an experiment under dynamic flow conditions is possible in many different ways, it depends on the type of results you aim to obtain:
        1. 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);
        2. 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.

      Payment Terms

      You can either order and pay via credit card or invoice (through a purchase order). Please note that some products are not available for direct purchase: just click "Request a Quote" on the product page and fill in the form to get more informations!

      Credit card 

      • Add your products to the cart
      • In the cart page, click on Proceed to Checkout
      • Complete your credit card informations (don't worry, we are using Stripe, a PCI-certified program specialized in online transactions)
      • That's it! You will receive an order confirmation within a few minutes, and our team will send you the invoice.

      Invoice

      • Add your products to the cart
      • In the website header, click on Request a Quote
      • Fill in the required informations, then click on Submit Quote
      • Our team will send you a quote shortly
      • After receiving the quote, just send your purchase order at contact@darwin-microfluidics.com
      • Our team will complete your purchase and send you the invoice!

      If you wish to pay in USD, AUD or GBP, please specify it in your PO. We can add the converted value in the required currency to the quote if requested.

       

      Shipping

      The shipping charges are calculated based on the weight of the package and on the country of destination. We try to offer you the best shipping rates and services, shipping all our orders through DHL Express. Generally, your order reaches your lab in only 2 to 3 days after we send it, no matter the destination! 

      Please note that our shipping incoterms are DAP (Delivered At Place). Please contact us if you prefer using your own courrier account or have any question!

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