How to quickly adapt cell cultures to serum-free cultures?

Biotechnology Channel News: Many cell lineages tend to adapt to serum-free and protein-free cultures, while other cells that are too demanding for the environment are difficult to adapt to change, which requires a more specialized approach to adaptation.
and depending on the characteristics of the cell line, there are two ways to adapt cells to serum-free culturestyl.
method of continuous adaptation/sequennonation is the method of continuous isolation, and the other method is direct adaptation.
method of continuous adaptation/sequenquenncing isolation is continuous adaptation/sequenquenncing isolation.
a series of serum reduction steps to adapt cells to serum-free culturestyl.
this is a preferred method and does not create too bad changes in the cell culture environment.
Continuous adaptation/sequenary isolation - Method 1 enables cells in the original culture to go through the following stages continuously: stage 1:75% to supplement the serum-replenished culture or 25% serum-free culture to 2:50% to supplement the serum-free culture or 50% to serum-free The culture base stage 3:25% supplemented with serum or 75% serum-free culturer stage 4:100% serum-free culturestyl continuous adaptation/sequenary isolation method - method 21, with the same concentration in the original culture, the serum is added to the serum-free culture.
the transfer of cells from the original culture medium containing serum to the serum-free medium, care should be taken to increase cell density in the new culture system.
allows cells to have an adaptation transition period.
2, according to method 1, slowly reduce the serum concentration, so that the cells at each stage have time to adapt.
3, once the serum addition is reduced to 0, the cell culture has several adaptation transition periods before it is used for analysis or other operations.
, another method of direct adaptation is direct adaptation.
, cells are transferred directly from serum-containing methyls to serum-free culturestyls.
large numbers of cells are transferred directly to new cultures that are serum-free or protein-free.
cells must be in the middle of exponential growth and have a survival rate of 90%.
about 50% of the base every 3 to 4 days to prevent it from becoming acidic.
maintain a higher-than-normal cell culture density until the cultured cells need to change the medium daily.
, the cells are amplification into multiple culture bottles.
is recommended during adaptation, cells are usually sensitive to PH and temperature changes.
allow cells to adapt to at least one transition period at each stage.
cell survival and cell density cells divide rapidly over long periods of time, and at the beginning of the adaptation process, the survival rate is greater than 90%.
this is essential.
when cells need to be passed on, it is recommended to use a small generation ratio of 1:2 or 1:3 to maintain higher cell density while providing cell growth factors that may contribute to cells during isolation.
the cell's transmission density is increased as it adapts to new culture conditions.
cell growth If the cell stops growing during the adaptation process, the cell needs to have more time to adapt to the culture base combination.
example, in stage 4 of Method 1, a small amount of serum (1-3%) was added to the serum-free methyl.
helps cells adapt more easily to new cultures.
then, during several transition periods, slowly reduce the amount of serum until the amount learned is 0.
serum-free and protein-free cultures of Meiatech, a suspended culture, do not contain the cell elongation shadows found in the serum.
during a transition period, the wall cells will begin to leave the surface and become suspended cell line to begin to grow.
cells may also change morphologically as a result of adaptation.
to monitor the growth rate of cells, as long as the cells can be integrated with the new system, any morphological changes are not important.
cell clumps tend to form during serum-free culture.
culture, gently break the clumps to disperse them.
transfer the culture from the stationary cell culture bottle to the rotating cone bottle, which may help prevent the formation of cell clumps.
because serum proteins tend to bind to antibiotics, it is not recommended to use them during serum-free adaptation.
if serum protein does not exist, antibiotic levels may be toxic to cells.
to ensure that a large number of cells with primary culturestyls are stored before starting the adaptation process.
cells do not survive to the next stage, the culture bottles used at each stage are preserved throughout the process.
.