Cell culture basics techniques and media essentially cell culture involves the distribution of cells in an artificial environment in vitro which is composed of the necessary nutrients ideal temperature gases ph and humidity to allow the cells to grow and proliferate.
Cell culture media protocol.
T175 30ml at 2e4 cells cm2.
Most cell lines can be grown using dmem culture media or rpmi culture media with 10 foetal bovine serum fbs 2 mm glutamine and antibiotics can be added if required see table below.
Then resuspend cells in sterile media to a suitable volume for counting.
Presence of serum in the media has many drawbacks and can lead to serious misinterpretations in immunological studies 2 3 a number of serum free media have been developed 4 5 these media are generally specifically formulated to support the culture of a single cell type such as knockout serum replacement and knockout dmem from thermo fisher scientific and mtesr1 medium from stem cell.
As the cells are in 100 ml media the next calculation is.
Based on count and viability data seed cell suspension for an appropriate flask and density e g.
Cell culture cell culture is one of the major tools used in cellular and molecular biology providing excellent model systems for studying the normal physiology and biochemistry of cells e g metabolic studies aging the effects of drugs and toxic compounds on the cells.
Basal media mem α provides the basic nutrients that cells need such as amino acids ions and a ph buffer.
In vivo when the study involves living biological entities within the organism.
2 cryopreservation 2 for cell culture.
2 a semi synthetic solid medium containing sucrose as c source and nitrate as the sole source of nitrogen useful for the general cultivation of fungi yeasts and soil bacteria.
Label culture flask with all necessary info e g.
Consult abcam counting cell using a hemocytometer protocol.
Check which culture media and culture supplements the cell line you are using requires before starting cultures.
100 ml x dilution factor.
This protocol is for making complete media suitable for culturing tenocytes tendon cells.
This will give a value for the volume of media the cells should be in.
9 cell culture antibiotic sterile filtered.
Add the correct amount of pre warmed culture media using serological pipette.
If the cell count is 55 x 10 4 ml and there is 100 ml of cell suspension.
It contains mem α 20 fbs 1 pen strep and a small amount of 2 mercaptoethanol.
