CULTIVATING VIRUSES WHY IS
THERE NO GROWTH?
Can you culture a virus?
what is needed to replicate?
How does it work?
HOW CAN YOU CULTIVATE VIRUSES?
(3)
WHAT ARE THE KINDS OF CELL CULTURES?
(3)
Suspension culture (not important for us now)
Primary Cell Culture
Monolayer culture
WHAT ARE THE KINDS OF CELL CULTURES?
Primary Cell Culture
what is it?
what is it good for?
what is it used for?
WHAT ARE THE KINDS OF CELL CULTURES?
Monolayer culture
explain
CELL CULTURE
How is this done?
Cell culture refers to the removal of cells from an animal or plant and their subsequent
growth in a favorable artificial environment.
Cell culture involves the growth of dispersed cells in-vitro, either as cells in
suspension, or as a monolayer on a solid surface such as a inner surface of polystyrene
culture flask.
CELL CULTURE
Suspension cultures
what is this?
Suspension cultures- Cells which do not
require attachment for growth or do not
attach to the surface of the culture
vessels. Can be propagated in suspension.
WHAT ARE THE KINDS OF CELL
CULTURES?
SUBCULTURE CELL CULTURES
explain
SUBCULTURE CELL CULTURES: THIS IS THE PASSAGE OF
CELLS FROM ONE CULTURE VESSEL TO ANOTHER IN ORDER
TO OBTAIN ADEQUATE FRESH NUTRIENTS AND SPACE.
* SUBCULTURES ARE REQUIRED FOR DEVELOPING CELL LINES
CELL LINES
what are the 2 kinds?
1-CONTINUOUS CELL LINES/IMMORTAL CELL LINES/HETEROPLOIDY CELL LINES
2-FINITE/DIPLOID CELL LINES
CELL LINES
what are these?
what can’t they be used for?
CELL LINES
what are they?
what can they be used for?
Types of cell cultures
Cell Line
Finite cell lines/Diploid Cell Lines
what kind of population?
what is the life span?
can they be subcultured? how?
where do they come from?
what do they retain ?
what 3 properties do they maintain?
what is the speed of growth?
are they easy or hard to use?
example?
**Homogenous **population of a single cell type – fewer cell types
Limited life span: May be sub-cultured up to 100 times before the cells die
Derived mainly from embryos; or from secondary cell cultures
Cell retains original morphology and diploid chromosome number
These cell lines exhibit the property of contact inhibition, density limitation and
anchorage dependence
The growth rate is slow and doubling time is around 24-96 hours
Technically, less hassle to use
Example: WI-38 (WI: Wistar Institute) is a diploid human cell culture line composed
of fibroblasts derived from lung tissue of a three month old white female fetus.
Types of cell cultures
Cell Line
Continuous cell lines/Immortal Cell Lines/Heteroploid Cell Lines
single or multiple cell type?
where are they derived from? (2)
limited or infinite cell division?
genetically normal or strange? why?
what is missing
speed of growth?
easy or difficult to use?
what are they prohibited from? why?
Cell cultures of a single cell type - Most homogeneous
Derived directly from cancer cells; or induced transformation of a primary or diploid
cell strain to divide indefinitely
Genetically weird – furthest from animal. Abnormal morphology & chromosome
number
Absence of contact inhibition and anchorage dependence
The growth rate is rapid and doubling time can be 12-24 hours
Hassle free to use
FDA prohibits their use in Vaccine Production Bluthey
are
cancerous
cells
Morphology of Cells in Culture
How many catagories?
what is used to catagorize them?
Cells in culture can be divided in to three basic categories based on their shape
and appearance
Morphology of Cells in Culture
what are the 3 catagories?
Morphology of Cells in Culture
Fibroblastic
explain
Morphology of Cells in Culture
Epithelial-like
explain
Epithelial-like cells are polygonal in shape with
more regular dimensions, and grow attached to a
substrate in discrete patches.
Morphology of Cells in Culture
Lymphoblast-like
explain
Lymphoblast-like cells are spherical in shape
and usually grown in suspension without attaching
to a surface.
CELL CULTURE ENVIRONMENT
CULTURE MEDIUM
what is it?
CULTURE MEDIUM: CONTAINS ALL THE NUTRIENTS REQUIRED FOR THE GROWTH
OF CELLS.
CELL CULTURE ENVIRONMENT
SERUM IN CULTURE MEDIUM
what is the most common?
what 3 things does it help do?
SERUM IN CULTURE MEDIUM:
* FETAL BOVINE SERUM IS THE MOST WIDELY USED SERUM IN CULTURE MEDIA
FOR GROWTH AND MAINTENANCE OF CELLS. IT AIDS IN THE CELL ADHESION,
REGULATION OF CELL MEMBRANE PERMEABILITY AND PROVIDING NUTRIENTS.
CELL CULTURE ENVIRONMENT
PHENOL RED PH INDICATOR
what does red mean
what does orange yellow mean
what does yellow mean
PHENOL RED PH INDICATOR IS THE INDICATION OF PH VIA COLOR CHANGE.
* IF THE SOLUTION IS RED THE PH IS >7 (BASIC)
* IF THE SOLUTION TURNS ORANGE/YELLOW THE PH IS <7 (ACIDIC)
”+” it changes
toyellow (this is your note, but it is too small forme to read. see slide 11)
CELL CULTURE ENVIRONMENT
CO2 LEVEL
relevance?
CO2 LEVEL: ENSURE TO MONITOR THE CO2 LEVEL WHEN USING MEDIA BUFFERED
WITH A CO2-BICARBONATE BASED BUFFER IN ORDER TO MAINTAIN THE PH OF THE
MEDIUM
CELL CULTURE ENVIRONMENT
ANTIMICROBIAL AGENTS
purpose?
ANTIMICROBIAL AGENTS: PREVENTS CONTAMINATION WITH BACTERIA,
MYCOPLASMA, YEAST, MOLDS, ETC
TRYPSIN
what is it? why is it used? what result?