10007123Cell Culturing
Course Information
Description
Covers the basic techniques of plant and animal cell culture. Plant unit includes media preparation isolation of explants and establishment of callus from suspension cultures, growth factor bioassays, regeneration of whole plants from tissue and plant genetic engineering techniques. Mammalian cell unit includes media preparation, maintenance of cultured cells, transfection of cultured cells, cloning, monoclonal antibody production, and ELISA assays. 
Total Credits
3
Course Competencies
  1. Analyze the classification of plants and their structure
    Assessment Strategies
    Written Product, Exam, and/or Skill Demonstration in Lab
    Criteria
    Outline the characteristics of vascular and nonvascular plants
    Examine the basic plant life cycle
    Explore mitosis and meiosis
    Compare and contrast dicots and monocots
    Identify parts of the whole plant structure
    Identify reproductive parts of plant
    Explain the basic structure and function of leaves, stems, and roots

  2. Maintain plant cultures with no contamination
    Assessment Strategies
    Written Product, Exam, and/or Skill Demonstration in Lab
    Criteria
    Explain the function of each media component
    Explore how plant hormones affect plant growth and organogenesis
    Describe how to prepare media aseptically
    Perform a bioassay for cytokinin activity
    Calculate and plot plant cell growth by determining root and shoot growth
    Dissect and culture parts and embryonic tissue of plants using bright-field microscopy and open-bench aseptic technique
    Outline, establish and maintain callus, meristem, and plant cell suspension culture
    Explain the basic causes of explant loss

  3. Examine genetic transformation of plants
    Assessment Strategies
    Written Product, Exam, and/or Skill Demonstration in Lab
    Criteria
    Explain the methods used for recombinant DNA transfer with plans including the use of Agrobacterium, electroporation and microprojectile bombardment
    Examine a basic technique for isolating DNA from plant material
    Demonstrate Agrobacterium-mediated transformation of plant tissue cultures

  4. Analyze the historical, present, and future aspects of animal cell culture
    Assessment Strategies
    Written Product, Exam, and/or Skill Demonstration in Lab
    Criteria
    Identify the major historic advances in cell culture
    Explain the advantages and disadvantages of cell culture
    Summarize the different types of cultures
    Examine the anatomy and function of subcellular organelles
    Examine the elements of selection in the origin and evolution of cell lines
    Explain the cell cycle and the signals from the environment that regulate it

  5. Implement laboratory safety and biohazard procedures
    Assessment Strategies
    Written Product, Exam, and/or Skill Demonstration in Lab
    Criteria
    Identify typical general safety hazards in a tissue culture laboratory
    Demonstrate an effective level of protection for various biohazard procedures
    Demonstrate effective disposal procedures for tissue culture materials

  6. Determine the equipment needs of an animal cell culture lab
    Assessment Strategies
    Written Product, Exam, and/or Skill Demonstration in Lab
    Criteria
    Confirm essential and beneficial equipment for a cell culture facility
    Identify other types of equipment that may be found in a cell culture facility
    Determine the necessary consumable items used in a tissue cell facility

  7. Demonstrate aseptic technique
    Assessment Strategies
    Written Product, Exam, and/or Skill Demonstration in Lab
    Criteria
    Describe the objectives of aseptic technique using biological safety cabinets (BSC)
    Demonstrate good technique for work surface, personal hygiene, pipetting, and sterile handling
    Explain the mechanism of biological safety cabinets (BSC)

  8. Prepare media for culturing cells
    Assessment Strategies
    Written Product, Exam, and/or Skill Demonstration in Lab
    Criteria
    Compare the surfaces and the dishes, plates, and vessels that cells will grow on
    Explain the relationship between CO2, temperature, buffering, and pH
    Explain the basic constituents of media
    Contrast the advantages and disadvantages of serum-free media
    Clean and sterilize equipment
    Prepare media in biological safety cabinets (BSC)
    Prepare media and sterilize by filtration
    Test media for sterility

  9. Maintain and passage cultured mammalian cells
    Assessment Strategies
    Written Product, Exam, and/or Skill Demonstration in Lab
    Criteria
    Culture CHO-KI, iPS (IMR90)-4, and 40-1a cell lines without contamination in biological safety cabinets (BSC)
    Culture cells in various kinds of incubation vessels
    Count cells using a hemocytometer
    Count cells using an automated cell counter (Countess)
    Calculate and prepare dilutions of cells
    Plate cells at a specific cell density
    Demonstrate cryopreservation techniques by freezing and thawing CHO-KI and iPS (IMR90)-4 cells

  10. Apply methods for manipulating and analyzing cultured cells
    Assessment Strategies
    Written Product, Exam, and/or Skill Demonstration in Lab
    Criteria
    Explain morphology and health of cell cultures using bright-field microscopy
    Describe methods for DNA, RNA and protein analysis of harvested cells
    Demonstrate how cell cultures can be used to assay viability, growth, and cytotoxicity
    Transfect CHO-KI cells with plasmid DNA
    Select and clone transfected cells
    Document the expression of recombinant protein in transfected cells using fluorescent microscopy
    Examine the basic biology of B cells and T cells
    Diagram an antibody molecule
    Compare polyclonal and monoclonal antibodies
    Examine the methods and uses for antibodies in ELISA, dot-blot, Western Blot, and hybridoma production

  11. Explain industrial aspects of cell culturing
    Assessment Strategies
    Written Product, Exam, and/or Skill Demonstration in Lab
    Criteria
    Describe industrial applications such as biopharmaceutical production, vaccine development, drug discovery, and food and beverage production
    Identify various products from cultured animal cells, such as the production of monoclonal antibodies, viral vaccines, or fermented foods
    Identify the basic components of a bioreactor

  12. Analyze the production and use of induced pluripotent stem cells in medical and research applications
    Assessment Strategies
    Written Product, Exam, and/or Skill Demonstration in Lab
    Criteria
    Compare the morphology of somatic and stem cells using bright-field microscopy
    Investigate current methods for the maintenance and use of induced pluripotent stem cells (IMR90-4)
    Explain differentiation methods and states for stem cell cultures