Course | Madison College Outline of Instruction

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

Course Competencies

Examine the classification of plants

Assessment Strategies

through the completion of written assignments

through written examinations

Criteria

you outline the characteristics of vascular and nonvascular plants

you examine the basic plant life cycle

you explore mitosis and meiosis

you outline the characteristics of gymnosperms and angiosperms

you compare and contrast dicots and monocots
Summarize the basic plant structures

Assessment Strategies

through the completion of written assignments

through written examinations

Criteria

you identify parts of the whole plant structure

you identify reproductive parts of plant

you explain the basic structure and function of leaves, stems, and roots
Examine the basic elements of plant culturing

Assessment Strategies

through the completion of written assignments

through written examinations

Criteria

you outline the time line for successful plant cell culturing

you outline the different methods of plant propagation

you explain the different types of plant growth

you explain the basic causes of explant loss
Examine components needed for media.

Assessment Strategies

through the completion of written assignments

through written examinations

Criteria

you summarize components needed for media

you explain the function of each media component

you summarize the plant hormones used to stimulate plant growth

you explore how plant hormones affect plant growth and organogenesis

you examine how to prepare media
Demonstrate the effects of hormones and chemicals on plant cultures.

Assessment Strategies

through the completion of laboratory activities

through the completion of written explanations of results and observations of laboratory activities

Criteria

you produce root and stem cultures from callus cultures with no contamination

you perform a bioassay for cytokinin activity

you calculate and plot plant cell growth by determining cell density using a hemacytometer
Establish and maintain plant cultures with no contamination.

Assessment Strategies

through the completion of laboratory activities

through the completion of written explanations of results and observations of laboratory activities

Criteria

you produce callus growth with two or more types of explants

you establish and maintain plant cell suspension culture

you establish and maintain a meristem culture
Manipulate plant protoplasts.

Assessment Strategies

through the completion of laboratory activities

through the completion of written explanations of results and observations of laboratory activities

Criteria

you isolate plant protoplasts from flower petals

you determine protoplast viability

you perform protoplast fusions to produce hybrid cells

you examine applications of plant protoplasts
Examine genetic transformation of plants.

Assessment Strategies

through the completion of written assignments

through the completion of laboratory activities

through written examinations

through the completion of written explanations of results and observations of laboratory activities

Criteria

you explain the methods used for recombinant DNA transfer with plans including the use of Agrobacterium, electroporation and microprojectile bombardment

you examine a basic technique for isolating DNA from plant material

you demonstrate Agrobacterium-mediated transformation of plant tissue cultures
Examine somaclonal variation and mutagenesis.

Assessment Strategies

through the completion of written assignments

through the completion of laboratory activities

through written examinations

through the completion of written explanations of results and observations of laboratory activities

Criteria

you examine the problems and advantages of genetic variation

you demonstrate the mutagenic properties of sodium azide on plant cells

you examine traits of crop plants studied for modification

you examine novel trait which could improve a plant variety
Examine products made by plant culture methods.

Assessment Strategies

through the completion of written assignments

through the completion of laboratory activities

through the completion of written explanations of results and observations of laboratory activities

Criteria

you explain the relationship between secondary plant metabolites and products of economic importance

you summarize the types of cultures used to facilitate production

you analyze the relationship between growth rate and production of secondary metabolites
Demonstrate the production and release of secondary matabolites by plant cells.

Assessment Strategies

through the completion of laboratory activities

through the completion of written explanations of results and observations of laboratory activities

Criteria

you use a cell culture to produce a secondary metabolite that inhibits the growth of algae or fungus

you demonstrate the use of a plant-derived biocatalyst
Examine the history of animal cell culture.

Assessment Strategies

through the completion of written assignments

through written examinations

Criteria

you identify the major historic advances in cell culture

you explain the advantages and disadvantages of cell culture

you summarize the different types of cultures
Examine the biology of the cultured cell.

Assessment Strategies

through the completion of written assignments

through written examinations

Criteria

you examine the anatomy and function of subcellular organelles

you examine the elements of selection in the origin and evolution of cell lines

you explain the cell cycle and the signals from the environment that regulate it
Examine laboratory safety and biohazard issues.

Assessment Strategies

through the completion of written assignments

through written examinations

Criteria

you outline the elements of risk assessment

you summarize the typical general safety hazards in a tissue culture laboratory

you explain the proper level of protection for various biohazard procedures

you explain proper disposal procedures for tissue culture materials
Examine the equipment needs of a tissue culture lab.

Assessment Strategies

through the completion of written assignments

through written examinations

through the completion of written explanations of results and observations of laboratory activities

Criteria

you examine essential and beneficial equipment for a tissue culture facility

you examine other types of equipment that may be found in a tissue culture facility

you examine the necessary consumable items used in a tissue culture facility
Examine aseptic technique.

Assessment Strategies

through the completion of written assignments

through the completion of laboratory activities

through written examinations

through the completion of written explanations of results and observations of laboratory activities

Criteria

you examine the objectives of aseptic technique

you examine good technique for work surface, personal hygiene, pipetting and sterile handling

you explain the mechanism of laminar flow hoods
Examine the factors that influence the growth of cells during incubation.

Assessment Strategies

through the completion of written assignments

through written examinations

Criteria

you compare the surfaces and the dishes, plates, and vessels that cells will grow on

you explain the relationship between CO, temperature, buffering, and pH

you explain the basic constituents of media

you contrast the advantages and disadvantages of serum-free media
Prepare media for culturing cells.

Assessment Strategies

through the completion of laboratory activities

through the completion of written explanations of results and observations of laboratory activities

Criteria

you clean and sterilize equipment

you prepare media and sterilize by filtration

you test media for sterility
Aseptically maintain and passage cultured cells.

Assessment Strategies

through the completion of laboratory activities

through the completion of written explanations of results and observations of laboratory activities

Criteria

you culture CHO-KI, NIH3T3, and NS-I cell lines without contamination

you establish a primary cell culture

you count cells using a hemocytometer

you demonstrate cryopreservation techniques by freezing and thawing CHO-KI cells
Manipulate transfected and transformed cultured cells.

Assessment Strategies

through the completion of laboratory activities

through the completion of written explanations of results and observations of laboratory activities

Criteria

you transfect CHO-KI cells with plasmid DNA

you select and clone transfected cells

you transform NIH3T3 cells and stains foci with Giemsa stain
Examine methods for analysis of cultured cells.

Assessment Strategies

through the completion of written assignments

through written examinations

through the completion of written explanations of results and observations of laboratory activities

Criteria

you examine methods for DNA, RNA and protein analysis of harvested cells

you explain how cell cultures can be used to assay viability and cytotoxicity
Examine industrial aspects of cell culturing.

Assessment Strategies

through the completion of written assignments

through written examinations

Criteria

you examine the basic components of a bioreactor

you summarize the concept of downstream processing

you examine various products from cultured animal cells
Examine basic immunology

Assessment Strategies

through the completion of written assignments

through written examinations

through the completion of written explanations of results and observations of laboratory activities

Criteria

you examine the basic biology of B cells and T cells

you diagram an antibody molecule

you compare polyclonal and monoclonal antibodies

you examine the methods and uses for antibodies, ELISA, Western Blot, and hybridoma production
Examine the use of animals in cell cultures

Assessment Strategies

through the completion of written assignments

through written examinations

through the completion of written explanation of results and observations of laboratory activities

Criteria

you examine the ethical considerations of animal use

you examine the regulations regarding animal care and use

you demonstrate proper handling and care of laboratory animals

you examine the proper injection technique for immunization of mice
Produce hybridomas with spleen from immunized mice and NS-I cells

Assessment Strategies

through the completion of laboratory activities

through the completion of written explanations of results and observations of laboratory activities

Criteria

you perform the fusion process to create hybridoma cells

you perform ELISA to screen hybridoma cells for antibody production