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Definition
Obtained from tissues of freshly dissected animals
Advantages: normal cells
Disadvantages: finite number of cell divisions, sometimes hard to obtain a “pure”/homogenous cell culture. |
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Term
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Definition
Primary cultures are allowed to divide
Advantages: cell differentiation can be studied. Ex: fibroblasts
Disadvantages: finite number of cell divisions, cell isolation process can potentially eliminate factors (cell-cell interactions) needed for cell differentiation. |
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Term
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Definition
Transformed cells, immortal
Advantages: homogeneity, techniques to produce mutants have been developed, can be used to produce recombinant proteins
Disadvantages: chromosomal abnormalities, malignancy |
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Term
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Definition
Used for biological product manufacturing
Original cell lines are obtained from reputable sources
produced from a single pool of cells, produced from a single clone
growing at standard, defined conditions
Genetically stable |
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Term
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Definition
Used for large-scale cell or tissue culture
Conditions such as: nutrient and oxygen delivery, waste disposal, pH, pressure, and temperature are highly controlled.
due to the large density of cells living in a 3D structure, Controlling appropriate nutrient delivery, physical stress, waste disposal are difficult |
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Term
| Process of extracting primary culture cells |
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Definition
Dissection
Removal of ECM by: Trypsin, collagenase, EDTA
Isolating wanted cell population using:
Centrifugation
Affinity binding to Ab
Binding to plastic surfaces
FACS
microdissection |
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Term
| Using FACS to isolate cells |
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Definition
Monoclonal antibodies, labeled with a fluorescent marker, are used against cell surface markers.
Cells bound to the mAb are sorted away from the mixture of cells and collected for further use. |
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Term
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Definition
Exploit the use of hybrid cell construction and media selection.
A normal B cell is fused with a myeloma cell line with PEG |
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Term
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Definition
| PEG causes the cells to fuse their cell membranes |
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Term
| Hybrid cells (such as b-cell and myeloma cells) are cultured in : |
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Definition
| The hybrid cell is cultured in HAT (hypoxanthine-aminopterin-thymidine) media |
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Term
| Myeloma cells can not grow on HAT media because they lack a functional: |
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Definition
| HGPRT gene. Aminopterin in media blocks new nucleotide synthesis. |
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