Cell Culture Flashcards
What are the requirements for cell culture?
- Sample of tissue
- method to digest tissue to a suspension of cells using proteases like trypsin to break cell-cell contacts
- Sterile conditions
- Tissue culture median. a growth liquid containing amino acids, vitamins, and glucose (energy source).
5.Blood serum which contains growth factors to support growth of cells - known growth factor supplement
7.antibiotics to keep down contamination - Surface to grow cells on usually a petri-dish – normal cells (eg Keratinocytes) require attachment
- Incubator kept at 37 c, humid and set to keep medium at pH 7.0. Controlled oxygen levels.
Explain skin grafting
Skin grafting - Burns – Can take the skin from the arm, leg or some other part of the body not affected by the burn, grow them in cell culture to build up the numbers, then graft the cells onto the wound site where they continue to grow
Uses of cell cultures
Amniocentesis - checking developing foetus for genetic abnormalities.
Carcinogen testing
IVF
Cloning
What conditions can be tested for using amniocentesis in a cell culture and what they cause
Down’s syndrome - trisomy of chromosome 21 - learning difficulties
Klinefelter’s Syndrome - male has XXY chromosomes instead of XY - sterile male, small besties, learning difficulties
Turner’s Syndrome - Sterile female, does not menstruate, short in stature, can have learning difficulties
How do you form a chromosomal spread
Cultured cells are incubated with colchicine (a drug which inhibits mitotic spindle) leading to mitotic arrest.
What is Geimsa
A stain which stains DNA blue/purple allowing abnormal chromosomes to be identified in a chromosomal spread
What are the benefits of 3D cultures over 2D
Tissue-Like Structure: 3D cell cultures mimic the three-dimensional architecture of tissues and organs more closely
Cell-Cell Interactions: Cells in 3D cultures have more opportunities for direct cell-cell interactions
Co-culture of multiple cell types
Diffusion gradient of drug and nutrients
