Analyzing Cell, Molecules and Systems 1 Flashcards
(45 cards)
What is the purpose of cell culture?
Study of cells, tissues, or organs in vitro. Also mimics the behaviour of the environment from where the cells were isolated
What does cell culture refer to?
Refers to the removal of cells from an organism, and promote their subsequent growth in a favorable artificial environment
Where are primary cell cultures derived from?
From an explant, directly from the animal
Cultured either as tissue explants or single cells;
Can be embryo or adult; normal or neoplastic
How long does the primary cell culture usually survive?
Only for a finite period of time
What does primary cell culture involve?
Enzymatic and/or mechanical disruption of the tissue and some selection steps to isolate the cells of interest from a heterogeneous population
What is the main difference between primary and cell line cultures?
Primary: transient
Cell lines: permanent
What are continuous cell lines?
A primary culture that has become immortal due to some transformation; most commonly tumour derived, or transformed with a virus such as Epstein-Barr
What are examples of Continuous cell lines?
CHO, SH-SY-5Y, Hela, K562, HEK293
How many times does a primary cell culture divide?
Divides only a limited number of times before losing their ability to proliferate
What is senescense?
Cells divide only a limited number of times before losing the ability to proliferate
Cell lines that have senescence are known as what?
Finite
What do primary cell cultures start out as?
A heterogeneous population
Cell line cultures are derived from what?
A primary or secondary culture
How are cell line cultures immortalized?
Spontaneous genetic mutation or by transformation vectors (viruses and/or plasmids)
What type of population are cell line cultures?
homogeneous
Cell line cultures will have ______ phenotype
Differentiated
What is the life span of cell line cultures?
Infinite life span in vitro, easy to grow and to cryopreserve for future experiments
Immortalied cell lines are isolated from naturally occurring cancer, major examples include:
human HeLa cells and mouse murine leukaemia cells
Mammalian cells in culture can be divided into 3 basic categories based on their morphology: describe
- Fibroblastic cells: bipolar or multipolar, have elongated shapes and grow attached to a substrate
- Epithelial-like cells:; polygonal in shape with more regular dimensions, and grow attached to a substrate in discrete patches
- Lymphoblast-like cells: are spherical in shape and usually grown in suspension without attaching to a surface
What are the advantages of cell cultures?
- study of cell behaviour without the variations that occur in animals
- Characteristics of cells can be maintained over several generations, leading to good reproducibility between experiments
- Control of the growth environment leads to uniformity of sample
- Cultures can be exposed to reagents
What are the disadvantages of cell cultures?
Have to develop standardized techniques in order to maintain healthy reproducible cells for experiments
Takes time to learn aseptic technique
Quantity of material is limited
Dedifferentiation and selection can occur and many of the original cellular mechanisms can be lost
It can be costly
What are the applications of cell cultures?
Basic research on cell/ gene function
Production of biological products (hormones, proteins, antibodies)
Testing of drugs, vaccines, chemical toxicity
Chromosomal or genetic analysis - clinical diagnostics
Regenerative medicine
What is hybridoma technology used for?
The generation of immortalized antibody-producing B cell lines, where an antibody-producing B cell is fused with a myeloma B cell cancer cell.
What is purification crucial for?
To study the structure and function of individual proteins
