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    3D Cell Cultures: Ajisafe Victor Ayobami

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    Ajisafe Victor Ayobami
    3D cell culture involves growing cells in all three dimensions to mimic their natural environment. This allows cells to interact with each other and their surroundings more accurately than 2D cultures. There are various techniques for 3D cell culture, including scaffold-based methods using polymeric scaffolds or biological materials, and non-scaffold methods like hanging drop plates, spheroid plates, microfluidics, and gels. These techniques are useful for applications in tissue engineering, drug testing, and cancer research.

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    3D Cell Cultures: Ajisafe Victor Ayobami

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    Ajisafe Victor Ayobami
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    3D cell culture involves growing cells in all three dimensions to mimic their natural environment. This allows cells to interact with each other and their surroundings more accurately than 2D cultures. There are various techniques for 3D cell culture, including scaffold-based methods using polymeric scaffolds or biological materials, and non-scaffold methods like hanging drop plates, spheroid plates, microfluidics, and gels. These techniques are useful for applications in tissue engineering, drug testing, and cancer research.

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    3D Cell Cultures

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    3D Cell Cultures

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    3D cell culture involves growing cells in all three dimensions to mimic their natural environment. This allows cells to interact with each other and their surroundings more accurately than 2D cultures. There are various techniques for 3D cell culture, including scaffold-based methods using polymeric scaffolds or biological materials, and non-scaffold methods like hanging drop plates, spheroid plates, microfluidics, and gels. These techniques are useful for applications in tissue engineering, drug testing, and cancer research.

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    72 views16 pages

    3D Cell Cultures: Ajisafe Victor Ayobami

    Uploaded by

    Ajisafe Victor Ayobami
    3D cell culture involves growing cells in all three dimensions to mimic their natural environment. This allows cells to interact with each other and their surroundings more accurately than 2D cultures. There are various techniques for 3D cell culture, including scaffold-based methods using polymeric scaffolds or biological materials, and non-scaffold methods like hanging drop plates, spheroid plates, microfluidics, and gels. These techniques are useful for applications in tissue engineering, drug testing, and cancer research.

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    of 16

    3D CELL CULTURES

    By:

    Ajisafe Victor Ayobami


    OSMANIA UNIVERSITY,
    HYDERABAD , INDIA
    What is 3D Cell Culture?
    • An artificially-created
    environment in which
    biological cells are permitted to
    grow or interact with their
    surroundings in all three
    dimensions.

    • An improvement over the


    previous method of
    growing cells in 2D.

    • Accurately mimics the cell


    natural environment.
    Why 3D Cell Culture?
    Before (2D) Now (3D)
    • Cells are grown on flat dishes and • Cells attach to one another and
    cellular attachment is unnatural. form cell-to-cell attachments
    • Cellular communication and synthesized by the cells.
    signaling is minimal. • Cellular communication and
    • Grown cells are simple with signaling is maximized.
    divergent functions from native • Grown cells have complex
    tissues. phenotype and functions that
    • Not valid for targeting and testing closely mimics native tissues
    new drug discoveries. • More valid for targeting and
    • Monolayer of cells are formed testing new drug discoveries.
    • Multi layers of cells are formed.
    2D vs 3D Cell Cultures
    3D Cell Culture Techniques
    a) Scaffold Based Scaffold Based
     Polymeric Hard Scaffolds
     Biologic Scaffolds
     Micropatterned Surface
    Microplates

    b) Non Scaffold Based


     Hanging Drop Microplates
     Spheroid Microplates
    containing Ultra-Low
    Attachment (ULA) coating
     Microfluidic 3D cell culture Non Scaffold Based

    3D Cell culture Bioreactor


    c) Bioreactors

    a) Gels
    3D Cell Culture Gels
    Scaffold Based
    Polymeric Hard Scaffold
    • Typical size is 150 - 200µm
    • Lactide and glycolide copolymer
    may be used as materials
    • In regenerative medicine, cells
    are grown on this scaffold for
    transplantation purposes to
    replace defective tissue
    • Widely used for engineering
    bone, cartilage, ligament, skin,
    vascular, neural and skeletal
    muscle tissues.
    • In preclinical in-vitro testing, cells
    are grown to model tumors in the
    lab.
    Scaffold Based
    Biological Scaffolds
    • Produced from natural
    components of biological origin
    e.g proteins such as fibronectin,
    collagen, laminin and gelatin.
    • Cells attach and reorganize into
    desired organs’ 3D structure.
    • Provides suitable
    microenvironment for the
    anatomical and physiological
    development of cells.
    • Widely used in regenerative
    medicine as well as tissue
    engineering
    Scaffold Based
    Micropatterned Surface Microplates
    • Each plate contains
    micrometer sized
    compartments arrayed on the
    bottom.
    • Usually coated to create a low
    adhesion surface.
    • Bottom is thin and transparent
    for microscopic imaging.
    • Used in drug testing, cell Round Square Slit pattern
    biology analysis at the
    molecular level
    Non Scaffold Based
    Hanging Drop Microplates
    • Cells can self assemble into 3D
    spheroid structure in the
    absence of a scaffold.
    • Spheroid size is controlled by
    the number of cells dispensed
    into each drop.
    • Suitable for long term culturing
    by transfer of culture onto
    another plate of larger media Top view Bottom view
    or buffer volumes
    • Suitable for use as primary cell-
    based tissue models and tumor
    models that include
    immortalized cancer cell lines
    Non Scaffold Based
    Spheroid Microplate
    • Used to create round multi
    cell tissue or tumor models.

    • Wells are shaped to a depth


    of SBS 96 – 384-well
    microplate.

    • Widely used for experimental


    purposes.

    • ULA is used to coat the


    bottom to minimize cell
    adherence for spheroid
    formation
    Non Scaffold Based
    Microfluidic
    • Used to create 3D architecture of in-vivo
    tissues and tumors
    • To create heterogeneous models.
    • Permits the continuously supply necessary
    nutrients and growth requirements as well
    as waste removal through culture medium.
    • Contains physical barriers that are made up
    of glass or silicon, polymers such as
    polycarbonate (PC), polystyrene (PS), and
    chromatographic or filter paper.
    • Cells may be combined with matrix to
    encourage cell-ECM interaction for the
    assembly of 3D structures.
    • Suitable for stem cells, primary and cancer
    cells
    Bioreactor
    • Ideal for mass cell production and ex vivo
    tissue engineering applications.
    • Suited for cell expansion applications or
    scaled production of cellular products, such
    as antibodies.
    • Bioreactors for 3D cell culture fall into 4
    main categories:
    – The spinner flask
    – Low-shear-stress rotating wall vessels
    – Perfusion bioreactors
    – Combination of bioreactors with
    scaffolds,
    Gels
    • Have soft-tissue like stiffness that
    mimic the ECM.
    • Can be made from ECM mixtures
    of natural sources such as
    collagen and alginate.
    • Most widely used is Matrigel
    • Synthetic gels include
    polyethylene glycol (PEG) – Based
    Hydrogel.
    • Difficult to use due to inability to
    maintain viscosity, and may be
    used in combination with other
    methods.
    Applications of 3D Cell Cultures
    Applications
    Consideration in 3D Cell Cultures
    CONCLUSSION
    • There are many methods
    available to create in-vivo
    cellular environments
    • Some are amenable for
    re-creating in-vitro barrier
    models.
    • others are more suitable
    for mimicking cancerous
    tumors or tissues.

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