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UTS KSO, 4 Kriteria Kultur Primer Cell Jelasskan yang dimaksud dengan…

- - - - Description: A small part of an organ is cultured on a filter or grid at a gas–liquid interface (usually with medium below the tissue).
      - Purpose: To maintain the original histological structure and cell interactions of the tissue.
      - Example: Culturing a piece of trachea or kidney tissue to study organ-level function.
      - Key Point: The 3D structure and cellular arrangement are preserved.
    - - Description: A small piece of tissue (explant) is placed at the solid–liquid interface (on a surface with medium partially covering it).
      - Behavior: Cells migrate out from the tissue fragment onto the surface, forming an outgrowth.
      - Use: Common for studying cell migration, wound healing, and tissue repair.
      - Key Point: Some tissue structure remains, but cells start spreading out.
    - - Description: Tissue is enzymatically or mechanically disaggregated into individual cells, which are then cultured as a monolayer.
      - Types:
        
        Primary culture: direct from tissue.
        
        Subculture: passaged cells forming continuous cell lines.
      - Use: For detailed studies of cell physiology, biochemistry, or drug testing.
      - Key Point: 3D structure lost, but cells are uniform and easier to manipulate.
      - Mechanical : high yield, faster,simple less expensive, use if tissue are plenty, risk os cell damage
        Enzymatic (Tripsin, kolagenase)
    - - Description: Cells from different sources (e.g., epithelial + stromal cells) are co-cultured on a support, sometimes with a matrix (e.g., collagen).
      - Purpose: To recreate a tissue-like structure in vitro.
      - Use: Modeling complex tissues (like skin or brain slices) to study cell–cell interactions or drug responses.
      - Key Point: Mimics tissue organization and function, bridging the gap between simple cell cultures and real tissues.
  - - - Culture medium, pH value (7,2-7,4), Effect of temperature (37C), Incubation (CO2 5%), Cryopreservation, Thawing, Passaging of cells, Trypsinization, Cell suspension, Cell count and cell infection, Cell harvest, Antibiotics, Laboratory management, Challenges during passage of cell, Maintenance of cell
- - - - cel to cell contact:
      - 2D Cell Culture
        
        flat shaped, not representative, easier to manipulate and analyze
      - 3D Cell Culture
        
        similar like in vivo, representative, types Scaffold free or scaffold based system, more physiologically relevant
        environment
        
        Scaffold-free system
        
        Spheroids Technique
        
        Organoids Technique
        
        Sacffold-based systems
        
        Solid scaffold
        
        Hydrogel
        
        Polymer-based
        
        Decellularized Tissue
        
        Microfluids
    - - autocrine or paracrine
        
        without direct contact whereas cytokine diffusion between containers is allowed
        
        porous membrane coculture (versatile tool)
      - process of obtaining Conditioned Medium (CM) a common method used to study cell-to-cell communication without direct contact between cells.
        
        Commonly used cell source MSC (Mesenchymal stem cell) dan Induced Pluripoten Stem Cell (iPSc) -> bone marrow, umbilical cord, wharton's jelly, Amnion-derived MSC
        
        How Condition media is produced
        
        Cell Isolation: collect from tissue
        
        Expansion: grow in cultur dish
        
        Collection: collect medium it has secreted molecule
        
        Storage: filter, clean stored
        
        This CM is then used to treat recipient cells, transferring the bioactive signals without the donor and recipient cells ever touching.
      - Generating CD-ECM (Cell-Derived Extracellular Matrix) fibroblas like cell used for scaffold or coating for another cell to grow on
        
        How CD-ECM is produced
        -Cell seeding-> Choose cell type, density,culture format
        
        ECM production-> Use low serum,vitamin, special condition to produce
        
        Decellularization: remove cell using detergent, keep ECM intact
        
        Processing: ECM can be freeze-dried, powder or made soluble
        
        Format of CD-ECM
        
        2D Culture: coating surface n easy to decellularize
        
        Standart: normal ECM
        
        On grooves: aligned ECM
        
        Crowded: dense ECM
        
        3D culture: mimic native tissue n allow complex shape
        
        Cell sheet/spheroids/Scaffold
        
        thicker, more realistic ecm structure
      - Coculture mediu optimization
        
        Culture Medium composition
        
        Culture Medium Volume
        
        Dialysis principle