Cell culture will be called as cell line after the first passaging of culture from a

primary cell culture. Generally, cell passaging or cell subculturing is the process that
involved transferring of cells from the previous medium to a new fresh medium to
prevent any death of cells causes by the accumulation of toxic materials,
insufficient of nutrients and increasing in cell numbers. Passaging of cells is
important to maintain a continuous growth of the cells with lower density of cells in
new medium, healthy cells colonies by providing fresh medium for continual growth
and reducing the possibility of cell death. There are several important requirements
that need to be considered when passaging cell lines which are the genome stability
of cells and the composition of the new medium. The composition of the new
medium must be same as the previous medium when passaging cell culture.
Adaptation of the cells towards the new medium will occur if different composition
are used during cell passaging. Moreover, multi-well culture plates from same
manufacturers must use during cell passaging because different manufacturer will
use different surface treatment to produce the culture well with a good adherent
surface or substratum for anchorage dependent cells. All of these might lead to
many genetic changes in the cells genome causes by the cells survival instinct.
(Masters & Stacey, 2007)
In this experiment, passaging or subculturing the monolayer of midgut cells had
been done in multi well culture plate. Two main types of the chemicals are used
during transferring of cells which are phosphate-buffered saline (PBS) and trypsin
EDTA. (Rakesh Arya., 2016) The main function of the PBS and trypsin EDTA are to
remove the serum of medium which can inactive the trypsin EDTA and to cut away
the focal adhensions that are anchoring the cell to the culture dish respectively. In
cell passaging, the optimal temperature for trypsin is 37 degree Celsius and it
normally will be used with EDTA that can help to remove trypsin inhibitor, calcium.
(Charles, 2016) The complete medium, DMEM will also be added later to stop or
deactivate trypsin EDTA.