Serum in Cell Culture - Use of FBS etc. in cell culture (Feb/06/2002 )
Does anyone have information on the effect of heat inactivation (56ºC for 30 min) on FBS.
Freshney (Culture of Animal Cells) states that it removes complement and reduces cytotoxic action of of immunoglobulins WITHOUT damaging growth factors. However I can't find any actual studies on this.
I am particularly interested in whether it effects connective tissue cells. Most papers mentioning heat inactivation are on immune cells.
I've looked at cell proliferation and seen little difference, but there do seem to be differences in certain phenotypic markers
in the Lab I work in, all FBS is heat inactivated in advance before use. One told me that e.g. ELISAs are not affected by the Complement System anymore once you inactivated the FBS.
I don't know any studies on that, but I can tell you, that my cell culture of HUMAN FIBROBLASTS work perfectly with inactivated FBS. So I guess the growth factors remain working.
Sorry, that I couldn't help more...
Kathrin, how long does it take for you to grow your cells?
I did a study recently and what took an avg of 6 to 7 days of confluence took only 3 days for mine.
it takes my fibroblasts at least 7 days until they reache confluence if I split them in a 1:2 ratio at passageing.
Since I work with patient materials I experienced that the speed of growth varies from patient to patient (sometimes it takes even 14 days).
Is it possible that your cell line turned tumorous? Are you working with normal fibroblasts? And how do you isolate your cells from tissue?