Animal biotechnology

Created by

Pierre Gasly

Cards (36)

What are the requirements to culture human cells
You require a supplemented growth media with vitamins, salts, glucose and essential amino acids.
Fetal bovine serum is used to provide the cells with growth factors.
Sterile conditions prevent competition with bacteria.
pH and Osmolarity are stabilised using phosphate buffered saline.
The cells are grown on plastic or glass surfaces.
What is confluence
The measurement of the percentage visual field covered by cells.
This is measured using a light microscope.
Label the cell stages in the tissue culture graph.
X.
A) Primary cell line
B) Transformed cell line
C) Senescence
What are primary cultures
These are the cells established directly from the tissue sample.
How are primary cell cultures made
Proteases such as trypsin digest the extracellular matrix and the cell to cell interactions.
Which type of cells secrete their own extracellular matrix
Fibroblasts
What are the stages in animal tissue culture
Primary culture
Primary cell line
Cell transfer
Senescence
Transformed cell line
What is a primary cell line
A monolayer of cells that descend directly from the tissue sample.
What is the role of porous membrane in animal culture
It mimics the basement membrane
How are cells transfered between dishes
Using trypsin
What is senescence
Cells can only grow and divide for a finite number of times usually 50 doublings. Beyond this point they will die.
What type of non-transformed cell can be cultured indefinitely
Embryonic stem cells
What is a cell line
These are a culture of cells with an indefinite life span. They have become transformed by spontaneous genetic changes.
What are the features of transformed cell lines
Immortal.
Anchorage independent.
Reduced requirement for growth factors.
Motile.
No contact inhibition of growth.
Tumourigenic.
Less differentiated.
What are HeLa cells
These are an immortalised cell line derived from cervical carcinoma.
What are polyclonal antibodies
They are produced from a mixture of B cell clones that recognise different epitopes
What are monoclonal antibodies
They are produced from a specific B cell that recognises a single epitope.
What is an antigen epitope
An amino acid sequence found on the antigen that cells recognize.
What are the key features of myeloma cells
These are transformed lymphocytes that do not produce antibodies.
How are monoclonal antibodies made
The mouse is injected with an antigen and this stimulates antibody production.
A spleen sample [contains B cells] is fused with a myeloma to form hybrids known as hybridomas.
Hybridomas are selected by culturing on a selective media.
Screen the library of hybridomas for the required antibody.
Culture the required hybridomas.
How are monoclonal antibodies used in research
They can be used as probes to detect proteins.
They can be used to purify antigens using affinity chromatography.
How are monoclonal antibodies used in diagnosis
They can be used to detect disease markers
How are monoclonal anitbodies used as a therapeutic treatment
Herceptin is the commercial name for trastuzumab which is used to treat HER breast cancer.
What is tissue engineering
The production of a functional construct to replace damaged tissue.
Describe how an ear reconstruction works
A scaffold is made from the mould of a normal ear. It is made of biodegradable material.
A biopsy takes a cartilage sample from the patient. These cells are isolated and expanded.
Seed the scaffold with the cells and allow them to grow.
Implants the cartilage graft into the patient.
What are the main properties of stem cells
They can divide and renew themselves.
They produce precursor cells which can form specialised cells.
What are the classes of stem cells
Pluripotent.
Totipotents.
Adult.
What are pluripotent stem cells
They have the ability to form all types of cells
What are adult stem cells
They are used for cell regeneration or tissue repair, and so only become certain cells.
What are pluripotent stem cells
They can divide infinitely and form all types of cells.
What are totipotent stem cells
They can form all types of cells
What type of stem cells are embryonic
These are pluripotent
Where are embryonic stem cells isolated from
The inner cell mass
How are iPS cells made  
4 stem cell master regulator genes are inserted into a retroviral vector which is inserted into a fibroblast.
This turned on the stem cell genes and form an iPS cell.
What is direct reprogramming
When a somatic cell type is reprogrammed into another.
How does direct reprogramming wor
You can make a transgene with a transcription factor and clone using a viral vector.
Change the endogenous gene expression using the CRISPR CAS 9 system. This allows you to cut the DNA and insert your required gene within.