Cell Culture Protocol - Essay Example

All movements within the cabinet should be slow, with no rapid movement, as slow movements help the air to circulate properly in the cabinet (MSUM, n.d.).3) Prepare a suitable culture medium (MSUM, n.d.).4) Check the pH of the culture medium by using the indicator phenol red (MSUM, n.d.).

Normally, cells can survive better in a slightly acidic medium than in alkaline media (MSUM, n.d.).5) Check for cell confluency and attachment (MSUM, n.d.) by comparing the occupied area with the unoccupied area (MSUM, n.d.).6) The floating cells should be dividing cells (MSUM, n.d.), and they should not be round unless they are dividing (MSUM, n.d.). The number of giant cells should be low and fixed (MSUM, n.d.).7) Discard the cell medium (MSUM, n.d.).8) Wash the cell layer with 1-2 ml of ‘calcium-magnesium free PBS’ (MSUM, n.d.).9) Pipette trypsin onto the cell layer (MSUM, n.d.) and incubate for 2-10 min to detach cells (MSUM, n.d.).10) Suspend the cells in fresh serum to inactivate trypsin (MSUM, n.d.).11) Titrate the cells to remove clumps (MSUM, n.d.), and shift the required number of cells into a new flask containing a fresh and pre-warmed medium (MSUM, n.d.).

The cell line selected for the study is rat coronary artery adventitial fibroblasts (Jenkins et al., 2007). The advantage of this selection is the fact that these cells have an important role in developing arteriosclerosis and plaque development (Jenkins et al., 2007), they divide rapidly and can differentiate into myofibroblasts (Jenkins et al., 2007) so those cells and their role in cardiovascular diseases can also be studied by the same culture instead of culturing the cells separately.