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Questions and Answers
What is the term for unorganized plant cells that can be grown in aggregated tissue masses?
What is the term for unorganized plant cells that can be grown in aggregated tissue masses?
Callus
What type of liquid media is used for cell suspension cultures?
What type of liquid media is used for cell suspension cultures?
Agitated liquid media
Cell suspension cultures are typically started by placing an inoculum of callus in a solid medium.
Cell suspension cultures are typically started by placing an inoculum of callus in a solid medium.
False
Under agitation, single cells in suspension culture break off and form cell chains and clumps.
Under agitation, single cells in suspension culture break off and form cell chains and clumps.
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It is always necessary to have a previous callus phase before initiating suspension cultures.
It is always necessary to have a previous callus phase before initiating suspension cultures.
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What is the name of the medium used for leaf sections in suspension culture?
What is the name of the medium used for leaf sections in suspension culture?
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What is the name of the structure that helps plant cells adhere to each other?
What is the name of the structure that helps plant cells adhere to each other?
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Cultures of completely isolated plant cells have been successfully obtained.
Cultures of completely isolated plant cells have been successfully obtained.
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What factors affect the proportion and size of cell aggregates in suspension cultures?
What factors affect the proportion and size of cell aggregates in suspension cultures?
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Cells divide more frequently in isolation than in aggregates.
Cells divide more frequently in isolation than in aggregates.
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The size of cell clusters increases during the phase of rapid cell division.
The size of cell clusters increases during the phase of rapid cell division.
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Agitation in batch cultures causes the size of cell clusters to increase as they approach a stationary growth phase.
Agitation in batch cultures causes the size of cell clusters to increase as they approach a stationary growth phase.
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What factor primarily influences cell dispersion in suspension cultures?
What factor primarily influences cell dispersion in suspension cultures?
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What type of growth regulators increase the specific activity of enzymes that dissolve the middle lamella of plant cell walls?
What type of growth regulators increase the specific activity of enzymes that dissolve the middle lamella of plant cell walls?
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To increase cell dispersion, a high concentration of auxin and a high concentration of cytokinin are used in the culture medium.
To increase cell dispersion, a high concentration of auxin and a high concentration of cytokinin are used in the culture medium.
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Using high auxin levels to achieve maximum cell dispersion ensures that the cultured cells remain differentiated.
Using high auxin levels to achieve maximum cell dispersion ensures that the cultured cells remain differentiated.
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Using suspension cultures to produce secondary metabolites is advantageous when high auxin levels are used.
Using suspension cultures to produce secondary metabolites is advantageous when high auxin levels are used.
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Which of these is NOT a type of cell suspension culture method?
Which of these is NOT a type of cell suspension culture method?
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What is the key characteristic of batch cultures?
What is the key characteristic of batch cultures?
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What is the key characteristic of continuous cultures?
What is the key characteristic of continuous cultures?
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What is the key characteristic of immobilized cell cultures?
What is the key characteristic of immobilized cell cultures?
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All cell suspension culture techniques utilize some method of agitating the culture medium.
All cell suspension culture techniques utilize some method of agitating the culture medium.
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Batch cultures are often used for large-scale industrial production of plant cells.
Batch cultures are often used for large-scale industrial production of plant cells.
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Continuous cultures are used for the large-scale production of primary or secondary metabolites.
Continuous cultures are used for the large-scale production of primary or secondary metabolites.
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Continuous culture techniques are simple and require minimal equipment.
Continuous culture techniques are simple and require minimal equipment.
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Mechanically stirred reactors can be used for large-scale suspension cultures without damaging plant cells.
Mechanically stirred reactors can be used for large-scale suspension cultures without damaging plant cells.
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The growth of plant cells is generally slower in suspension than in callus cultures.
The growth of plant cells is generally slower in suspension than in callus cultures.
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Root and shoot initiation in cell suspensions usually commence in isolated single cells.
Root and shoot initiation in cell suspensions usually commence in isolated single cells.
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Somatic embryos can only arise from callus tissues.
Somatic embryos can only arise from callus tissues.
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Cells from suspension cultures can be plated onto solid media to regenerate plants.
Cells from suspension cultures can be plated onto solid media to regenerate plants.
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It is not possible to obtain plants from somatic embryos formed in suspension cultures.
It is not possible to obtain plants from somatic embryos formed in suspension cultures.
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What are the traditional methods of cell suspension culture assessment?
What are the traditional methods of cell suspension culture assessment?
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What are the two main techniques used to assess cell viability in suspension cultures?
What are the two main techniques used to assess cell viability in suspension cultures?
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What is the key takeaway message from today's lecture?
What is the key takeaway message from today's lecture?
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Study Notes
Cell Suspension Culture in Plant Tissue Culture
- Plant cells in suspension culture can be grown as callus in aggregated tissue masses or freely dispersed in agitated liquid media.
- Techniques for cell suspension culture are similar to those used for large-scale bacterial cultures.
- Cell suspension cultures are initiated by introducing a friable callus inoculum into a liquid medium.
Lesson Objectives
- Students will understand the concept and significance of cell suspension culture.
- Students will learn the process of establishing and maintaining cell suspensions.
- Students will discuss the applications and advantages of cell suspension culture.
Cell Suspension Culture
- Under agitation, single cells break off and form chains or clumps.
- Subsequent fracturing creates individual cells or small cell groups.
- A previous callus phase is not always necessary for starting suspension cultures.
- Leaf sections placed on MS medium and exposed to light show rapid growth and cell division in the mesophyll.
- After four days on a rotary shaker, leaf sections can be disintegrated to release a large number of cells into suspension.
Cell Suspension Culture (continued)
- Plant cell walls adhere naturally, hindering the creation of suspensions containing only single cells.
- Cultures with fully isolated cells have yet to be achieved.
- The proportion and size of cell aggregates depend on the plant variety and growth medium.
- Cell clusters tend to increase in size during rapid cell division.
Cell Suspension Culture (continued)
- Agitation detaches single cells and small cell groups in batch cultures, reducing cluster size as cultures near a stationary growth phase.
- The degree of cell dispersal in suspension cultures is affected by growth regulator concentrations in the medium.
- Auxinic growth regulators increase enzyme activity, dissolving the middle lamella in plant cell walls.
- Using a high auxin concentration and a low cytokinin concentration in the medium often increases cell dispersal.
Cell Suspension Culture (continued)
- High auxin levels during cell dispersion maintain the cultured cells in an undifferentiated state.
- This can be a disadvantage when the suspension is used for producing secondary metabolites.
Cell Growth Curve
- The provided graph illustrates a cell growth curve with phases including lag, exponential growth, linear growth, stationary, and culture inviability.
Cell Suspension Culture Methods
- Batch cultures: Cells are grown in a fixed volume of media until growth ceases.
- Continuous cultures: Cell growth is maintained by continuous replenishment of sterile nutrient media.
- Immobilized cell cultures: Plant cells are immobilized in inert matrices, with fresh media constantly supplied.
Additional Notes on Cell Suspension Culture
- The growth of plant cells in suspension culture is typically more rapid and controllable compared to callus cultures.
- Culture media can be easily altered in suspension cultures.
- Root and shoot initiation often begins in cell aggregates.
- Somatic embryos can develop from single cells.
Additional Notes on Cell Suspension Culture (continued)
- Plants can be grown from somatic embryos formed in suspension cultures.
- Embryos are developed into plantlets using solid media.
- In suspension cultures, single cells or cell aggregates grow onto solid media and form callus colonies.
- Regenerated plants can develop from these cultures.
Assessment of Cells in Suspension Culture
- Cell count
- Packed cell volume
- Cell fresh weight
- Optical density
- Viability test (using fluorescein diacetate (FDA) and Evans blue stains)
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Description
This quiz explores the techniques and processes of cell suspension culture in plant tissue cultures. Students will learn about the initiation, maintenance, and applications of these cultures, as well as their significance in research and agriculture. A solid understanding of these concepts will enhance students' knowledge in tissue culture methodologies.
