Cell Encapsulation Platform

    A cell encapsulation platform inside double emulsions

    The cell encapsulation platform, developed and manufactured by Secoya and constituted by Fluigent’s flow control equipment and Secoya's Emulsion Technology, complete system for high throughput encapsulation of complex and individual cells within highly monodisperse double emulsion droplets small enough (<90 µm) for further analysis.

    This fast and easy system allows the encapsulation of cells in aqueous core (such as medium, PBS) and oil shell (such as HFE 7500) double emulsions, providing a powerful tool for biochemical and cellular assays, as it enables the isolation of each cell within microreactors, highlighting its characteristics and concentrating the signals to measurable levels to obtain meaningful biological data.

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    Cell encapsulation platform for double emulsion in microfluidics developed by Secoya
    Main benefits
    • droplet monodispersity
      High precision

      Down to 2% monodispersity

    • ICO_BFT size
      Flexible

      Adjustable droplet size and shell thickness

    • ICO_BFT continuous
      Robust

      Continuous production 

    • Easy to use

      Intuitive interface and ready-to-use system

    • Biocompatible
      High Encapsulation efficiency

      Up to 100%

    Features of the cell encapsulation platform

    ➤ Fluigent Precision and Flexibility

    Produce robust and highly monodisperse emulsions using Fluigent’s pressure-based flow controllers and the Raydrop while accurately controlling the size of your droplets and shell thickness.

    ➤ Start emulsions production right away

    The system is a fully equipped, mounted and controlled tool to create double emulsions with little setup time.

    ➤ Complete and easy to use engineered system

    Our organized system requires easy priming and cleaning processes for better performance. It includes dedicated optics for optimized droplet visualization at high frequency.

    ➤ Innovative and much sought-after application for cell analysis

    The system is an easy-to-use platform for creating single-cell encapsulations into double emulsion droplets compatible with high-throughput screening.

    The cell encapsulation platform allows:

    • Elimination of the risk of cross-contamination.
    • Fast and efficient mixing of the reagents that occurs inside droplets.
    • Ability to work with cells of limited availability.

    Description

    The cell encapsulation platform includes an organized flow path with pressure controllers, filters, flowmeters, and valves to facilitate start-up, shut-down, and cleaning of the system between runs. Dedicated optics are included to optimize the visualization of droplet production at high generation frequency.

    Raydrop Double Emulsion Device

    RayDrop Double Emulsion

    Microfluidic Double Emulsion Device

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    LINK Microfluidic Software control

    LINK

    Microfluidic Software control

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    P-CAP series

    Air-tight tube metal cap

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    FLOW UNIT microfluidic flow sensor

    FLOW UNIT | FLOW UNIT +

    Bidirectional Microfluidic Flow Sensor

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    Flow EZ™ microfluidic flow controller

    Flow EZ™

    Microfluidic flow controller

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    Cell encapsulation platform: Examples of use

    Standard cell encapsulation platform

    The standard cell encapsulation platform allows the encapsulation of single cells for high throughput screening applications involving drug discovery, toxicity testing, various ‘omics’ studies and rare cell analysis1,2.

    Fig 1. Standard cell encapsulation platform.

    How to inject a small sample?

    The cell encapsulation platform with an injection loop allows working with samples with a reduced volume (e.g. working with cells of limited availability, such as stem cells, primary cells from patients…). Ideal for the use of rare cells or expensive reagents with a total volume lower than 500µl/.

    Fluigent L Switch

    L-SWITCH™ 6-port/2-position

    Microfluidic Recirculation Valve

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    Cell encapsulation platform for double emulsion in microfluidics developed by Secoya
    Cell encapsulation platform set up
    Fig 2. Cell encapsulation platform with a injection loop (L-Switch).

    How to add a reagent?

    In many microfluidic procedures, an integrated tooling for mixing is essential for appropriate functionality in a broad range of applications.

    The mixing cell encapsulation platform allows to integrate a new phase for optimal agent mixing. It has been optimized to mix two solution the later in a fluidic path before encapsulation. After precise formation of self‐ assembled lipid or polymeric particles by microfluidic mixing, encapsulation of active molecules is loaded into the synthesized particles.

    The efficiency of many biosensors and the possibility of investigating reaction kinetics with good time resolution depend on controlled mixing. It can be also used to encapsulate cells with barcoded beads for Next generation sequencing (DropSeq), for enzymatic test or phenotyping5.

    Cell encapsulation system
    Fig 3. Mixing cell encapsulation platform with an additional Flow EZ and a Flow Unit M.
    FLOW UNIT microfluidic flow sensor

    FLOW UNIT | FLOW UNIT +

    Bidirectional Microfluidic Flow Sensor

    Read more
    Flow EZ™ microfluidic flow controller

    Flow EZ™

    Microfluidic flow controller

    Read more
    Cell encapsulation platform for double emulsion in microfluidics

    Cell encapsulation platform for the production of double emulsions compatible with FACS.

    The platform allows you to produce double emulsions with aqueous core and oil shell (water-oil-water) to encapsulate cells (from 5 µm to 40 µm) making it compatible with FACS for cell sorting applications.

    Cell encapsulation platform: Applications

    High-throughput screening: Cell sorting

    Cell encapsulation can improve high-throughput screening outcomes since the highly monodisperse droplets provide homogeneous reaction conditions. It can be combined with cell sorting tool such as Fluorescence Activated Cell Sorting (FACS), to achieve higher speed and efficiency compared to conventional approaches.

    • Lower costs while increasing sequencing accuracy and depth:
      • Sorting and sequencing only cell containing droplets could vastly reduce reagent costs and increase accuracy and depth for downstream Next Generation Sequencing, including eliminating common issues with single-cell droplet sequencing such as reads from empty droplets due to encapsulation of free-floating transcripts.
    • Increased range of measurement:
      • Encapsulated cells may be sorted based on phenotypes not currently measurable with standard cell sorting, such as enzymatic turnover, presence of secreted molecules, or quantification of proteins lacking cell surface markers.
    • Multi-omics profiling:
      • In contrast with conventional methods, cell sorting using microfluidics provides a unique tool to link genotype with phenotype through compartmentalization.
      • Our cell encapsulation platform allows the combination of single-cell phenotyping and genome-wide sequencing, enabling multiomic measurements and directly linking cellular phenotypes to their underlying genetic mechanism2.
    Cell encapsulation method
    Figure 4. Selection of Double Emulsion Microdroplets Using a Fluorescence-Activated Cell Sorter (FACS).
    E. Mastrobattista, V. Taly, E. Chanudet, P. Treacy, B. T. Kelly, A. D. Griffiths, Chemistry & Biology 2005, 12, 1291.

    Drug delivery

    Cell or drug encapsulation for drug delivery consists in the immobilization of bioactive materials, mainly cells, within a double emulsion, generally surrounded by a polymeric membrane. The latter permits the free passage of nutrients and oxygen and the egress of therapeutic protein products. Encapsulation allows the protection of the cell content from mechanical stress and in the case of allogeneic tissue also from host’s immune response. Microcapsules are commonly used in pharmaceutical or medical processes as drug carriers or for the encapsulation of organic cells

    Locally-implanted droplets may represent a minimally invasive manner of delivering therapeutic proteins such as growth factors and cytokines. It could also allow mosaic injections containing several droplets to provide complex release profiles or multi-protein delivery3.

    monodispersed Cell encapsulation platform
    Figure 5. Stable monodispersed microcapsules with a solid PLGA shell and an aqueous core.

    Transplants

    Microencapsulation of living cells may serve as an alternative therapy for patients requiring organ transplants. Researchers can control droplets size to mitigate the immune infiltration of transplanted cells while keeping oxygen and waste products transported. Droplets can also immobilize cells at the desired transplant size, essential since many cell therapies rely on systemic cell administration.

    Cell encapsulation for cell-based therapy is emerging as a promising strategy for treating a wide range of human diseases, such as diabetes, blood disorders, acute liver failure, spinal cord injury, and several types of cancer through transplants. Pancreatic islets, blood cells, hepatocytes, and stem cells are among the many cell types currently used for this strategy. The encapsulation of these “therapeutic” cells is intended to prevent immune rejection, to provide a controlled and supportive environment, and to enable a complete retrieval of the graft in the case of an adverse body reaction4.

    cell encapsulation in chitosan
    Figure 6. Chitosan-shell/oil-core microcapsules.

    Specifications

    We propose 4 different platforms in order to target different droplet sizes.

    DescriptionProductP.N
    Cell encapsulation deviceCell encapsulation platform O-FACS1-PTF
    Double Emulsion production device– RayDrop Double Emulsion 30-70-45
    		– O-DE-RDRPC05-
    EUP
    Fluid handling system– LINK Module
    – Flow EZ 7 bars for all 3 phases
    		– LU-LNK-0002
    – LU-FEZ-7000PCK
    Reservoirs
    – Continuous phase:
    50 mL Pcap with 50 mL Falcon tube
    Tubing: 500 µm
    – Shell phase:
    15 mL Pcap with 15 mL Falcon tube
    Tubing: 125 µm
    – Core phase:
    15 mL Pcap with 15 mL Falcon tube
    Tubing: 125 µm

    		– P-CAP50-HP-PCK
    – P-CAP15-HP-PCK
    – P-CAP15-HP-PCK
    Flow Meters

    – Continuous phase: Flow Unit M+
    – Shell phase: Flow Unit S
    – Core phase: Flow Unit M
    		– FLU-M+-PCK
    – FLU-M-D-8PCK
    – FLU-S-D-8PCK
    Optical System
    – Light source
    – Microscope objective (x10)
    – Specific colour camera (up to 400 fps, 1µs integration
    time)
    – XYZ translation stages
    		N/A
    Tubing & Fittings
    – Tubing:
    OD: 1/16 and 1/32 OD
    ID: 125 µm & 500 µm
    Materials: PEEK, PFA

    – Manual valves:
    4 way valves
    2 way valves

    – Filters:
    10 µm filter for continuous phase
    2 µm filter for dispersed phase
    		N/A
    Wetted materials
    – Platform: PEEK, PFA, PCTFE, PTFE, SS316L, GLASS
    – Sealing: FKM or EDPM
    		N/A
    Unit dimensions
    – 61 x 46 x 43cm 3 (L x W x H)
    		N/A
    Weight
    – 4 kg without the protective wood
    – 21 kg with the protective wood
    		N/A
    Droplet Size Range*
    25-45 µm
    		N/A
    * Formulation dependant
    DescriptionProductP.N
    Cell encapsulation deviceCell encapsulation platformO-FACS2-PTF
    Double Emulsion production device– RayDrop Double Emulsion 30-70-60
    		– O-DE-RDRPC06-
    EUP
    Fluid handling system– LINK Module
    – Flow EZ 7 bars for all 3 phases
    		– LU-LNK-0002
    – LU-FEZ-7000PCK
    Reservoirs
    – Continuous phase:
    50 mL Pcap with 50 mL Falcon tube
    Tubing: 500 µm
    – Shell phase:
    15 mL Pcap with 15 mL Falcon tube
    Tubing: 125 µm
    – Core phase:
    15 mL Pcap with 15 mL Falcon tube
    Tubing: 125 µm

    		– P-CAP50-HP-PCK
    – P-CAP15-HP-PCK
    – P-CAP15-HP-PCK
    Flow Meters

    – Continuous phase: Flow Unit M+
    – Shell phase: Flow Unit S
    – Core phase: Flow Unit M
    		– FLU-M+-PCK
    – FLU-M-D-8PCK
    – FLU-S-D-8PCK
    Optical System
    – Light source
    – Microscope objective (x10)
    – Specific colour camera (up to 400 fps, 1µs integration
    time)
    – XYZ translation stages
    		N/A
    Tubing & Fittings
    – Tubing:
    OD: 1/16 and 1/32 OD
    ID: 125 µm & 500 µm
    Materials: PEEK, PFA

    – Manual valves:
    4 way valves
    2 way valves

    – Filters:
    10 µm filter for continuous phase
    2 µm filter for dispersed phase
    		N/A
    Wetted materials
    – Platform: PEEK, PFA, PCTFE, PTFE, SS316L, GLASS
    – Sealing: FKM or EDPM
    		N/A
    Unit dimensions
    – 61 x 46 x 43cm 3 (L x W x H)
    		N/A
    Weight
    – 4 kg without the protective wood
    – 21 kg with the protective wood
    		N/A
    Droplet Size Range
    45-60 µm
    		N/A
    * Formulation dependant
    DescriptionProductP.N
    Cell encapsulation deviceCell encapsulation platformO-FACS3-PTF
    Double Emulsion production device– RayDrop Double Emulsion 60-120-60
    		– O-DE-RDRPC07-
    EUP
    Fluid handling system– LINK Module
    – Flow EZ 7 bars for all 3 phases
    		– LU-LNK-0002
    – LU-FEZ-7000PCK
    Reservoirs
    – Continuous phase:
    50 mL Pcap with 50 mL Falcon tube
    Tubing: 500 µm
    – Shell phase:
    15 mL Pcap with 15 mL Falcon tube
    Tubing: 125 µm
    – Core phase:
    15 mL Pcap with 15 mL Falcon tube
    Tubing: 125 µm

    		– P-CAP50-HP-PCK
    – P-CAP15-HP-PCK
    – P-CAP15-HP-PCK
    Flow Meters

    – Continuous phase: Flow Unit M+
    – Shell phase: Flow Unit S
    – Core phase: Flow Unit M
    		– FLU-M+-PCK
    – FLU-M-D-8PCK
    – FLU-S-D-8PCK
    Optical System
    – Light source
    – Microscope objective (x10)
    – Specific colour camera (up to 400 fps, 1µs integration
    time)
    – XYZ translation stages
    		N/A
    Tubing & Fittings
    – Tubing:
    OD: 1/16 and 1/32 OD
    ID: 125 µm & 500 µm
    Materials: PEEK, PFA

    – Manual valves:
    4 way valves
    2 way valves

    – Filters:
    10 µm filter for continuous phase
    2 µm filter for dispersed phase
    		N/A
    Wetted materials
    – Platform: PEEK, PFA, PCTFE, PTFE, SS316L, GLASS
    – Sealing: FKM or EDPM
    		N/A
    Unit dimensions
    – 61 x 46 x 43cm 3 (L x W x H)
    		N/A
    Weight
    – 4 kg without the protective wood
    – 21 kg with the protective wood
    		N/A
    Droplet Size Range
    50-60 µm
    		N/A
    * Formulation dependant
    DescriptionProductP.N
    Cell encapsulation deviceCell encapsulation platformO-FACS4-PTF
    Double Emulsion production device– RayDrop Double Emulsion 60-120-90
    		– O-DE-RDRPC08-
    EUP
    Fluid handling system– LINK Module
    – Flow EZ 7 bars for all 3 phases
    		– LU-LNK-0002
    – LU-FEZ-7000PCK
    Reservoirs
    – Continuous phase:
    50 mL Pcap with 50 mL Falcon tube
    Tubing: 500 µm
    – Shell phase:
    15 mL Pcap with 15 mL Falcon tube
    Tubing: 125 µm
    – Core phase:
    15 mL Pcap with 15 mL Falcon tube
    Tubing: 125 µm

    		– P-CAP50-HP-PCK
    – P-CAP15-HP-PCK
    – P-CAP15-HP-PCK
    Flow Meters

    – Continuous phase: Flow Unit M+
    – Shell phase: Flow Unit S
    – Core phase: Flow Unit M
    		– FLU-M+-PCK
    – FLU-M-D-8PCK
    – FLU-S-D-8PCK
    Optical System
    – Light source
    – Microscope objective (x10)
    – Specific colour camera (up to 400 fps, 1µs integration
    time)
    – XYZ translation stages
    		N/A
    Tubing & Fittings
    – Tubing:
    OD: 1/16 and 1/32 OD
    ID: 125 µm & 500 µm
    Materials: PEEK, PFA

    – Manual valves:
    4 way valves
    2 way valves

    – Filters:
    10 µm filter for continuous phase
    2 µm filter for dispersed phase
    		N/A
    Wetted materials
    – Platform: PEEK, PFA, PCTFE, PTFE, SS316L, GLASS
    – Sealing: FKM or EDPM
    		N/A
    Unit dimensions
    – 61 x 46 x 43cm 3 (L x W x H)
    		N/A
    Weight
    – 4 kg without the protective wood
    – 21 kg with the protective wood
    		N/A
    Droplet Size Range
    70-90 µm
    		N/A
    * Formulation dependant

    OxyGEN

    Control in real-time, protocol automation, data record and export
    ver. 1.0.0.0 or more recent

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    Software Development Kit

    Custom software application
    ver. 21.0.0.0 or more recent

    See the offer

    References

    1. Steele, J.A.M. et al. (2014) “Therapeutic cell encapsulation techniques and applications in diabetes,” Advanced Drug Delivery Reviews, 67-68, pp. 74–83. Available at: https://doi.org/10.1016/j.addr.2013.09.015.
    2. Collins, D.J. et al. (2015) “The Poisson distribution and beyond: Methods for microfluidic droplet production and single cell encapsulation,” Lab on a Chip, 15(17), pp. 3439–3459. Available at: https://doi.org/10.1039/c5lc00614g.
    3. Headen, D.M., García, J.R. and García, A.J. (2018) “Parallel droplet microfluidics for high throughput cell encapsulation and synthetic microgel generation,” Microsystems & Nanoengineering, 4(1). Available at: https://doi.org/10.1038/micronano.2017.76.
    4. Farina, M. et al. (2019) “Cell encapsulation: Overcoming barriers in cell transplantation in diabetes and beyond,” Advanced Drug Delivery Reviews, 139, pp. 92–115. Available at: https://doi.org/10.1016/j.addr.2018.04.018.
    5. Damiati, S. et al. (2018) “Microfluidic devices for drug delivery systems and drug screening,” Genes, 9(2), p. 103. Available at: https://doi.org/10.3390/genes9020103.

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