By Hitendra R.H. Patel, Manit Arya, Iqbal Shergill
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Additional info for Basic Science Techniques in Clinical Practice
Two types of cultured cell types: • Primary cells • Cell lines Primary Cells These are cells derived directly from tissue samples/biopsies which have heterogeneous nature of cells with variable growth friction. The cells are extracted directly from the tissue and grown directly in speciﬁed optimal cell culture Medias. Cell Lines These are cells subcultured from primary cells but now have been manipulated in the laboratory so that they last longer and can go through more cell passages with growth friction of 80% or more, than the original primary cells before they change their morphology.
This buffer contains a tracking dye optionally; reducing agents, such as dithiothreitol (DTT) or 2-mercaptoethanol; and, most importantly, SDS. Heat and reducing agents destroy tertiary protein folding and quaternary protein structures by breaking up disulﬁde bonds and preventing their reformation simultaneously. SDS is an anionic detergent that denatures secondary and nondisulﬁde-linked tertiary structures, thus unfolding and linearizing the proteins completely. 0 g protein. Hence, it applies a negative charge to each protein in proportion to its mass (number of amino acids) so that they may be separated in an electric ﬁeld strictly by their length or size, respectively.
If the cells remain uncontaminated, then they are returned back to a normal media for another 2 to 3 days to conﬁrm that no contamination remains. First-choice antibiotics/antifungal agents for cell culture: • • • • Gentamicin Streptomycin Penicillin G Amphotericin B (antifungal) 4. 1. Cell culturing in a laminar ﬂow cabinet. Note the scientist is using a battery-operated “pipette boy” to draw the media up into the pipette. N. ) Media: Their Function in Cell Culturing Media provide the food and supplements that are needed for the growth of cells.