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Lecture 1

BIOL 4004 Lecture Notes - Electric Field, Senescence, Western Blot

5 Pages
89 Views
Fall 2015

Department
Biology
Course Code
BIOL 4004
Professor
Matthes David
Lecture
1

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MBC5 Study Guide – Chapter 8 (Manipulating Proteins, DNA, and RNA)
Unit !When reading chapters !Read the text of chapter 8 including these figures
1!10 and 11 !8-2 to 8-11
2!12 and 13 !8-12 to 8-23
3!14 and 15 !8-24 to 8-30; 8-33, 8-51
4!16 and 19 !8-34 to 8-52; 8-55, 56
5!17 and 18 !8-66 to 8-75
6!20 and 23 !8-57 to 8-65
ISOLATING CELLS AND GROWING THEM IN CULTURE
A cell culture refers to a population of cells that have been grown in the laboratory. Cell
culturing methods enable cell biologists to obtain large numbers of cells for experimentation. In
addition, the components that are added to a cell culture can be controlled, which is another
experimental advantage. In this section, we will consider some of the issues associated with cell
culturing methods, and how cells can be manipulated experimentally.
Cells Can Be Isolated from intact Tissues
The cells that a cell biologist wishes to study may come from a variety of sources. If the cells are
derived from unicellular species, such as bacteria or yeast, such cells are typically obtained from
colleagues or scientific stock centers at various places around the world. If the cells are from a
multicellular species, such as animals and plants, the situation is a bit more complex. In some
cases, a researcher may begin with a tissue sample from an organism that contains more than one
type of cell. For example, a small tissue sample from the tail of a mouse would contain skin
cells, blood vessel cells, etc. To start a cell culture, researchers often times want that culture to be
composed of a single type of cell. Therefore, the different cells types within a tissue must be
separated from each other. Figures 8-2 and 8-3 describe two methods for separating individual
cells. You should be familiar with the first method, fluorescence activated cell sorting. The
second method shown in Figure 8-3 will not be covered in this course. You may read it for
interest.
Cells Can Be Grown in Culture
Certain types of cells will grow on solid growth media. The cells tend to adhere to that media. In
the case of animal cells, the solid media is often coated with a protein, such as collagen, that
enables the cells to adhere. One advantage of solid growth media is that it enables researchers to
visualize the cells as they are growing. We will consider methods of cell visualization in Chap 9.
Serum-free, Chemically Defined Media Permit Identification of Specific Growth Factors
Certain types of cells will grow in liquid media. Such liquid media can have a defined
composition, making it possible for researchers to identify factors that are necessary for cell
growth. Furthermore, the composition of the growth media can be altered to study its effects on
cell division, cell function, etc. For example, a hormone can be added to a liquid culture of cells,
and then a researcher can study the effects of that hormone on cell structure and/or function.
Eukaryotic Cell Lines Are a Widely Used Source of Homogeneous Cells
A cell line is a population of cells that have been derived from a single cell via many, many cell
divisions. Many cell lines that are used by researchers are immortal, which means that they will
continue to divide without becoming senescent. An immortal cell line is an experimental
advantage because researchers can continue to propagate more cells as long as they want. Some
immortal cell lines were originally derived from tumors. An example is the HeLa cell line.
Alternatively, other immortal cell lines were derived from normal tissues, but later incurred
mutations during growth in cell culture that made them immortal.
Some examples of cell lines are shown in Table 8-1. Please do not memorize this table. Cell lines
are shared among researchers. When scientists use the same cell lines in their experiments, it is
easier to directly compare each others results.
Embryonic Stem Cells could revolutionize medicine
Embryonic stem cells can proliferate indefinitely in culture and give rise to all cell types in the
body. Since ES cells can behave like normal cells, they could potentially be used for tissue
repair, for example to replace cardiac muscles after a heart attack or to replace dead nerve cells
in patients with Parkinson's disease. However, ES cells may also produce tumors called
teratomas or they may be rejected by the recipient's immune system.
Somatic Cell Nuclear Transplantation May Provide a Way to Generate Personalized Stem
Cells
Cells from adult tissues can be used to generate clones of genetically identical cells through a
technique called somatic cell nuclear transplantation. The nucleus of an unfertilized egg cell is
replaced by a nucleus from a somatic cell from an adult organism. Such hybrid cell can give rise
to an early embryo, which can be transferred to the uterus of a foster mother to generate a whole
new animal (reproductive cloning) or be used as a source of stem cells to treat diseases or for
tissue repair. These procedures, especially reproductive cloning, are controversial, and outlawed
in some countries.
In 2006, researchers found a way to reprogram cells from adult tissues to regain properties of
embryonic cells. These cells, known also as Induced Pluripotent Stem Cells (IPSC), could
become any cell type in an organism given the right conditions and differentiating factors and
thus could become a source of cells for therapy. Indeed, these cells were successful in
generating a cloned organism.
Hybridoma Cell Lines are factories that produce Monoclonal Antibodies.
In certain types of experiments described later in this course, different cells are made to fuse
with one another. This initially creates a heterokaryon, which is a cell with two nuclei. Later,
hybrid cells will be produced that have a single nucleus containing genetic material from both of
the original cells. If one of he parent cells was from a tumor cell line, the hybrid cell is called a
hybridoma. Hybridoma cell lines are used extensively to produce monoclonal antibodies which
recognize specific proteins.
PURIFYING PROTEINS

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Description
MBC5 Study Guide Chapter 8 (Manipulating Proteins, DNA, and RNA) UnitWhen reading chapters Read the text of chapter 8 including these figures 110 and 11 82 to 811 212 and 13 812 to 823 314 and 15 824 to 830; 833, 851 416 and 19 834 to 852; 855, 56 517 and 18 866 to 875 620 and 23 857 to 865 ISOLATING CELLS AND GROWING THEM IN CULTURE A cell culture refers to a population of cells that have been grown in the laboratory. Cell culturing methods enable cell biologists to obtain large numbers of cells for experimentation. In addition, the components that are added to a cell culture can be controlled, which is another experimental advantage. In this section, we will consider some of the issues associated with cell culturing methods, and how cells can be manipulated experimentally. Cells Can Be Isolated from intact Tissues The cells that a cell biologist wishes to study may come from a variety of sources. If the cells are derived from unicellular species, such as bacteria or yeast, such cells are typically obtained from colleagues or scientific stock centers at various places around the world. If the cells are from a multicellular species, such as animals and plants, the situation is a bit more complex. In some cases, a researcher may begin with a tissue sample from an organism that contains more than one type of cell. For example, a small tissue sample from the tail of a mouse would contain skin cells, blood vessel cells, etc. To start a cell culture, researchers often times want that culture to be composed of a single type of cell. Therefore, the different cells types within a tissue must be separated from each other. Figures 82 and 83 describe two methods for separating individual cells. You should be familiar with the first method, fluorescence activated cell sorting. The second method shown in Figure 83 will not be covered in this course. You may read it for interest. Cells Can Be Grown in Culture Certain types of cells will grow on solid growth media. The cells tend to adhere to that media. In the case of animal cells, the solid media is often coated with a protein, such as collagen, that enables the cells to adhere. One advantage of solid growth media is that it enables researchers to visualize the cells as they are growing. We will consider methods of cell visualization in Chap 9. Serumfree, Chemically Defined Media Permit Identification of Specific Growth Factors Certain types of cells will grow in liquid media. Such liquid media can have a defined composition, making it possible for researchers to identify factors that are necessary for cell growth. Furthermore, the composition of the growth media can be altered to study its effects on
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