several authors report cell survival for 48h upon glucose depletion , in my hands after 15hours...100% cells death..Hela, U2OS..any hints to share.!
In my experience glucose deprivation is highly cell specific. I've seen breast cancer lines that begin apoptosis within 24 hrs and pancreatic lines that persist for a week or more without apparent ill effect (as they can very efficiently use glutamine as a carbon source for both energy and raw material) and some primary cells that enter a quiessent state. I'd suggest determining the effects of glucose deprivation on a cell by cell basis.
thanks, yes i imagine cells responsiveness vary..in our case, we tried to reproduce the data from this nat comm paper "Regulation of energy homeostasis by the ubiquitin-independent REGg proteasome," mainly the higher sensitivity of KO PA28 cells to GD...without success despite being in the same hela or u2os cells. we are using dialysed FBS or not, the authors just email us the expt work..in our hands, two independant researchers ..cells are dying...after OVN culture , we are using crisp KO cells and the CTL are dying...so the mystery remains...is there anything wrong with the way we process the cells!
You may have already considered these things but here are my top three reasons for unusual proliferation/apoptosis results.
1. Mycoplasma contamination, it can have a big affect when cells are stressed
2. Compiment proteins present in serum, be sure you are heat inactivating if the original authors did
3. Phenol pH indicator in the media, this depends on the androgen receptor status of your cells but that pH indicator can act as an estrogen mimic.
Best of luck
This is a point to be studied...
It is sure that most of the researchers do not try glucose deprivation since they mostly believe in glucose depretion, a situation fairly different.
The experiments needed to be done by many orher laboratory to be sure and design a unified results.
But I feel that this is quite interesting... Is glucose replaced by another sugar i.e. galactose?
in PC 12 cells I used glucose-free medium to test the effects on cell survival, but I did not observe any cell death during a 48 h period, probably the cells may rely on glutamine for survival
merci dominique..nous avons donc un pb...dans nos mains, des basiques Hela ou u2os meurent des que on retire le DMEM/SVF/glucose. on cultive en DMEM/glutamine sans glucose auquels on ajoute du SVF dialysé (serum dit "SILAC" si on rajoute du SVF non dialysé, les cellules sont ok) et PSG
dans quoi cultivez vous vos PC12... on lave les cellules 2-3x et on ajoute simplement le milieu. ..16-19h apres on analyse ..mais les cellules flottent..
et ceci dans les mains de deux experimentateurs ...experimentés:-)
Nous avons utilisé du DMEM sans rouge de phénol, supplémenté avec du sérum de veau (10%) et du sérum de cheval (décomplémentés) et 2mM L-glutamine....
Vous pourriez aussi trouver quelques renseignements sur des études faites il y a longtemps sur des hybridomes murins D Duval et al. Factors controlling cell proliferation and antibody production in mouse hybridoma cells: 1 Influence of the aminoacid supply Biotechnol Bioeng 38 (1991) 561-570
ou encore
D Duval et al. comparison of various methods for monitoring hybridoma cell proliferation J Immunol Methods 41 (1990) 235-242
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