Immortalised Cell Lines Flashcards
1
Q
What are immortalised cells ?
A
- either animal/human sources grown in culture for long periods of time
- dervived from multiple sources & contain chromosomal abnormalities /mutations which allow them to divide repeatedly
2
Q
What are some advantages of using immortalised cells?
A
- grow easily compared to primary cells (more robust)
- rapid growth
- standardised cells used by different labs across the world, offers consistency
3
Q
What are some disadvantages of using cell lines
A
- genetically altered - not ‘normal’ cells
- after continuous growth for long time, cell characteristics change further
- can be contaminant of other cell cultures as they grow so well
4
Q
What was 1 of the first cell lines to be established?
A
- HeLa Cells
- sourced from patient with aggressive cervical cancer, Henrietta Lacks
- estimated that this cell line has contributed to saving around 10 million lives
5
Q
Describe the developement of new immortalised cell lines
A
- key to cell immortalisation is the impairment of ;
1. cell cycle checkpoints - p53/p16 & pRb
2. upregulation of telomerase enzyme
3. upregulation of some oncogenes & oncoproteins
6
Q
Describe Transfection
A
- plasmids carrying important genes can be created & grown in bacteria
- then isolated, purified & transfected into eukaryotic cells
- cells will then hopefully express the gene introduced
- mRNA transfection is a way of bypassing the nucleus & potential binding issues
7
Q
Describe 3T3
A
- mouse embryonic fibroblasts
-robust & easy to handle - contact inhibited
- stops growing at very high densities
8
Q
Describe COS
A
- monkey kidney
- efficiently transfected
- commonly used as an expression system for high-level, short-term expressions of proteins
9
Q
What cell line is used to study melanoma?
A
- MV3 - human melanoma cell line, developed 1991
- highly metastatic
- developed by transplanting parts of melanoma into nude mice
-tumours were excised & grown in culture
10
Q
Describe A549
A
- Human Lung cancer cell line
- developed from alveolar basal epithelial cells
- 58 yr old man was initial donor & line was started in 1972
11
Q
Describe K-562
A
- originates from a chronic myeloid leukemia patient aged 53
- isolated from bone marrow
- similar to stem cell as its partly undifferentiated
12
Q
What cell line is used to study bone cancers?
A
- osteosarcoma cells such as MG-63
- Established from osteosarcoma in 1977 from a white 14 year old boy
- have a fibroblast like morphology
13
Q
How is aseptic technique achieved in labs ?
A
- class II sterile laminar flow hoods
- cells cultured in media that has been prepared under sterile conditions
- any tools/material used in contact with cells are sterilised using an autoclave/cleaned with 70% ethanol or irriated to ensure sterlitity
14
Q
How do Class II sterile laminar flow hoods create a sterile environment?
A
- creating an environment that is free of bacterial by positive air flow
15
Q
What is needed for culture media to grow eukaryotic cells?
A
- water
- salts
- protein sources
- carbohydrate sources
-antibiotic/antifungal compounds - growth factors specific to that cell type
16
Q
Describe tissue culture flasks
A
- flasks are treated so the surface chemistry of the flask is the correct surface energy for cells to stick to the surface
- disposable plastic
17
Q
What is a primary cell?
A
- a cell extracted from a tissue that is grown without any adulteration/transfection
- usually grows & proliferates more slowly
- require more complex culture conditions
18
Q
Why can cell lines not be appropriate ?
A
- responses to materials differs wildly to those of normal cells
