Download presentation
Presentation is loading. Please wait.
1
Chapter 6 Manipulating Cells in Culture
2
Advantages of working with cultured cells over intact organisms More homogeneous than cells in tissues Can control experimental conditions Can isolate single cells to grow into a colony of genetically homogeneous clone cells
3
Growth of microorganisms in culture Examples: E. coli and the yeast S. cerevisiae Have rapid growth rate and simple nutritional requirements Can be grown on semisolid agar Mutant strains can be isolated by replica plating Yeast colonies
4
Growth of microorganisms in culture
5
Replica plating
6
Growth of animal cells in culture Requires rich media including essential amino acids, vitamins, salts, glucose, and serum Most grow only on special solid surfaces A single mouse cell Figure 6-36 A colony of human cellsMany colonies in a petri dish
7
Growth of animal cells in culture
8
Primary cells and cell lines Primary cell cultures are established from animal tissues Most cells removed from an animal grow and divide for a limited period of time (about 50 doublings), then eventually die Certain “transformed cells” may arise that are immortal and can be used to form a cell line Transformed cells may be derived from tumors or may arise spontaneously
9
Establishment of a cell culture Figure 6-37
10
Cell fusion Two different cells can be induced to fuse thereby creating a hybrid cell (heterokaryon) Interspecific hybrids may be used for somatic-cell genetics Certain hybrid cells (hybridomas) are used to produce monoclonal antibodies
11
De Novo and salvage pathways for nucleotide synthesis Figure 6-9
12
Figure 6-10 Producing a monoclonal antibody to protein X
13
Chapter 5.5 Purification of cells and their parts
14
Figure 5-34 Purification of specific cells by flow cytometry Requires fluorescent tag for desired cell
15
Example: FACS data Figure 5-35
16
Purification of cell parts Understanding the roles of each each cell component depends on methods to break open (lyse) cells and separate cell components for analysis Cell lysis is accomplished by various techniques: blender, sonication, tissue homogenizer, hypotonic solution Separation of cell components generally involves centrifugation
17
Cell fractionation by differential centrifugation Figure 5-36
18
Organelle separation by equilibrium density-gradient centrifugation Figure 5-37
Similar presentations
© 2025 SlidePlayer.com. Inc.
All rights reserved.