Week 4: Growing and Studying Neural Cells in a Dish Flashcards
The cultivation of eukaryotic tissues or dissociated cells outside of the organism, in a growth media with the necessary nutrients, inorganic salts, and pH required to function in a physiologically normal manner.
Tissue Culture
Culturing of dissociated cells rather than pieces of tissues.
Cell Culture
Why is tissue culture useful?
It’s useful as a model system for studying the basic processes of cell biology; also has many clinical applications.
What are some examples of tissue culture in a clinical application?
- Diagnosis of chromosomal disorders from the culture of blood or amniotic fluid samples.
- Generation of monoclonal antibodies for the production of vaccines as a result of the development of hybridoma cell lines.
- In vitro fertilization, through techniques developed for the culture of the early embryo.
Most cell types, with the exception of blood cells, grow attached to an _________ _________
Extracellular Matrix (ECM)
Composed of a complex mixture of polysaccharides and proteins such as collagens and laminin
The Extracellular Matrix
____ _______ ________ on the surface of many cells bind strongly to the components of the ECM. These contacts have to be disrupted to detach cells without causing cell death.
Cell adhesion molecules.
The use of _______ also allowed for the re-plating of cells grown attached to a substrate.
Trypsin
Dissociated cells can be frozen indefinitely in ______ _______
Liquid nitrogen
Most modern tissue culture vessels are made of _______, are _____, and are intended for ________ use.
-reduces the risk of microbial contamination and cross-contamination of cell lines
plastic; sterile; single
Most tissue culture is performed in plates or flasks in varying sizes, often in varying sizes to prevent what?
airborne particles entering while allowing the free exchange of gasses
Widespread use of the antibiotics, penicillin and streptomycin, in the 1940s + reduced the problem of ______ contamination of cultures?
Microbial
_______ & _______ are ineffective against certain common strains of bacteria, such as mycoplasma
Penicillin and Streptomycin
The smallest known bacterial cells, making them difficult to observe under conventional light microscopes (<1 micron in length)
Mycoplasma
What is the best practice to prevent contamination?
Employing an aseptic technique
What is an antibiotic that IS effective against mycoplasmas?
Ciprofloxacin
One of the most important developments in improving aseptic techniques, designed to protect the user from hazardous microbes
Biological safety cabinets
Protects both the user and the environment from the sample; does not protect the sample from airborne particles from the environment
Class I cabinets
Protects samples from outside contamination; relies on a continuous uniform flow of clean filtered air travelling down over the sample; effective @ reducing microbial contaminants
Class II Cabinets
Completely encloses the sample, which can be accessed through the gloves integrated into the cabinet
Class III cabinets
Tissue culture incubators maintain a number of critical parameters to allow optimal growth and survival, which are:
- Constant levels of temperature, humidity, CO2, and Oxygen
- Most media currently in use for tissue culture use buffers that require an atmosphere of 5% CO2 to maintain a physiological PH
- Some cell types s/a pluripotent stem cells grow better under low oxygen conditions - specific incubators can reduce oxygen levels w/ displacement by nitrogen
1961, Hayflick and Moorhead derived the first strains of human fibroblasts (WI-38) young human diploid cells, making the distinction between what?
primary cells, cell lines, and cell strains
Are derived from normal tissue and grown without passaging.
Primary cells
Are derived from primary cells which a limited capacity for growth and division, but retain a normal karyotype
Cell Strains
Have the capacity to grow indefinitely and invariably have abnormal karyotypes
Cell Lines
The size, shape, and number of chromosomes in a cell
Karyotype
In 1951, George Otto Gey cultured cells from Henrietta Lacks with cervical cancer, discovering that the cells derived from the cervical tumour could do what?
Grow and divide indefinitely
& generated a cell line from a single cell - HeLa cells
Aided the development of the first Polio vaccine
HeLa Cells
A major advance in the development of cell lines that can be grown indefinitely & yet retain a normal karyotype & also the capacity to differentiate into specific cell types
Embryonic stem cells
Established cultures of cells derived from mouse blastocysts can in principle generate:
any cell type of the body in a cell culture dish
The first generated human embryonic stem cells from human blastocysts allowed for the generation of:
Inaccessible cell types, such as neurons, in large numbers for the first time
Through a process called reprogramming, it is possible to produce what by directly manipulating somatic cells, such as fibroblasts?
Induced Pluripotent Stem Cells (IPSCs)
Used to study genetic diseases and inaccessible cell types, s/a neurons
IPSCs
Transplanted cells will be genetically identical to that individual, eliminating the risk of rejection or the use of immunosuppressive drugs - describes a major advantage of what type of medicine?
Regenerative Medicine
