Since I don't have the cell culture facilities, I am maintaining the cells by transplanting in mice, but I am facing problem to maintain the EAC cells. Does anybody have any recommendations?
Swiss albino mice inoculated with EAC cells can be comfortably maintained for 7-10 days. The important points to be noted are:
1) 0.2 ml of EAC cells to be inoculated intraperitoneally.
2) Viability to be confirmed by tryphan blue dye exclusion method.
3) Minimum 10 animals to be kept for maintenance.
4) Avoid multiple punctures while collection. Also avoid collecting cells alongwith blood (only peritoneal fluid should be present).
Hope this helps.
What kind of problem are you facing? Please give me some details. (How many cells do you inoculate into mice periton? What did happen after inoculation: there was no proliferation, mice died, etc.) May be I can tell something.
hai freinds, this is shyam sunder, Asst professor in pharmacology, andhrapradesh. its a good method for anticancer drug screening, especially the researchers who are not having their institutional cell culture lab. during my study i have not found any problems, before start your experiment u should take some things into consideration:
do your work in cool conditions that is dont perform in summer.
some mortality i have also observed, so that u increase the no. of animals upto 10 in each gp.
u should check the viability of EAC cells by trypan blue dye and inject 0.2ml by IP route to each animal that is almost equal to previous woks done on the same cells.
u should select an appropriate duration of work maximum of 9 days.
ok bye, all the best
@Asha:
Hi asha i got it from
AMALA cancer research institute , thrissur, Kerala
only on Tuesdays it will be given to the mice,
you have to take the mice from your place
Swiss albino mice inoculated with EAC cells can be comfortably maintained for 7-10 days. The important points to be noted are:
1) 0.2 ml of EAC cells to be inoculated intraperitoneally.
2) Viability to be confirmed by tryphan blue dye exclusion method.
3) Minimum 10 animals to be kept for maintenance.
4) Avoid multiple punctures while collection. Also avoid collecting cells alongwith blood (only peritoneal fluid should be present).
Hope this helps.
I´d like to know if any of you wich work with EAC have experienced any problems after a certain number of passages.
I received an aliquot of EAC from Unicamp - SP/ Brazil, and it was explained to me that we need to maintain until 20 passages (after 5-7 days of inoculation - reference: OZASLAN, 2011) from a animal to another, 1x10^6 cells/mL, injecting 0.2 mL in each animal, i.p. I did an adaptation, injecting 0.5 mL to each animal.
So, we need to have these cells in -80 oC freezer, so we can take them to new animals and, in theory, we make the cells "turn back to" first passage (before the ones we are using get to 20th passage). The problem is that I haven´t seen this (the renewal of the cells) in any paper. Because of that, I need to see other people´s experiences.
The problem that is happening now is in the experiments itself: the volum of ascites are growing more in the drug treated groups than in control ones!
So, I hope my experience helps, also hoping to see yours.
i did many study with EAC, but there is no issues with maintaining as well as study animals. can you tell me the problem what do you found with this cell line?. it may help us too.
I agree with @Monalisa Brito, even after injecting 70 million cells in one mice there was problem in obtaining a fluidic cancer inside the mice. Hence number of passages greater than 25 are not advisable. Maintaining in mice is easy but only when you have a continuous running culture. Suggestions for the peoples who are starting it to grow for the first time please.
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