Ho Chi Minh City Medicine and Pharmacy University
Question
Asked 2nd May, 2016
Culturing EOL-1 cell line (a human eosinophil cell line)?
Dear all,
I would like to ask for your advice with culturing culture Eol-1 cells (eosinophilic cell line). I resuscitated cells from frozen stock and suspended in RMPI 1640+ 10% FBS + 1% PS. After 2 days, cells began to form clusters (as in the image that I enclosed) so I pipetted them slowly to dissociate cells. On the next day, cells started gathering again and they seemed unhealthy. So, I don't know if I did something wrong. Should I pipette more frequently or should I change new media? Can I continue using these cells?
According to Sigma's instruction, EOL-1 cells grow slowly as large clusters. Cells should be left to grow and not agitated more than necessary (http://www.sigmaaldrich.com/catalog/product/sigma/94042252?lang=ko®ion=KR).
Can anyone please share your experience with EOL-1 cells? I am grateful for every advice that you may give.
Best regards,
Tu Trinh
Most recent answer
Hello Frances,
According to what we have known so far, the treatment time and the media conditions will depend on your target outcome.
Here are some references that I have found so far, and I hope it would be helpful to you
I will use RPMI-1640 plus 2% FBS and 1% antibiotics for the normal treatment of eosinophils. The treatment time is varied from 6 hours to 16 hours, however, eosinophils will undergo the cell death cycle so you need to be careful with that. I have never let my cells in the media for more than 24 hours.
In addition, we observe that the coculture of eosinophils with other types of cells can protect eosinophils survival, in that case, we just use the RPMI+1% antibiotics without serum.
Sorry for late reply!
Tu
All Answers (9)
German Cancer Research Center
Hi,
I don't have any experience with these specific cells but I think the answer is in the citation you gave from sigma. If they want to be left alone, don't disturb them. :)
1 Recommendation
Retired
"Once upon a time" ... when I had "significantly less gray hairs" ... we performed the experiments in the attached article. It was with eosinophils obtained directly from human volunteers. However, I guess that the culture conditions would be the same with a "line".
Best regards
Robert
1 Recommendation
Ho Chi Minh City Medicine and Pharmacy University
Dear Gensch,
Thank you for your reply. Honestly I am a newbie with cell culture experiments so I am worried about "not agitated more than necessary " :-). However, I tried comparing two conditions of culture: in one flask, cells were left alone totally and in another flask, I pipetted cells gently enough just to disrupt the large clusters. There was not much difference between two flasks and cells grew quickly after 2 days of subculture.
Sincerely,
Tu
Ho Chi Minh City Medicine and Pharmacy University
Dear Professor Kiss,
Thank you for your nice paper, it helped me to understand more about eosinophil experiment.
As in the article, eosinophils were cultured in RPMI. May I ask if you have ever seen clumps of eosinophils during culturing? If there was, can you share with me how to avoid it? Because I would like to treat cells with some drugs and I want to make sure that each single cell can be exposed well to drugs. I appreciate any help you may provide.
Sincerely yours,
Tu
Retired
Dear Tu,
I am sorry but the expriments of this article were performed a long time ago and I do not remember the details.
In addition I was not personally culturing the eosinophils.
It seems to me that we did not face this problem of clumping because we were monitoring single eosinophil migration.
Sorry to be of so poor help.
Best regards
Robert
1 Recommendation
School of Pharmacy Medical Sciences Campus
Hello!
Have anyone of you any standard protocol to culture eosinophils? I will be working with eosinophils isolated from peripheral blood from humans and I would like to know how to manage them. For example, the time for subculturing and also the appropriate media conditions form them.
Any information will be really useful.
Thanks
Ho Chi Minh City Medicine and Pharmacy University
Hello Frances,
According to what we have known so far, the treatment time and the media conditions will depend on your target outcome.
Here are some references that I have found so far, and I hope it would be helpful to you
I will use RPMI-1640 plus 2% FBS and 1% antibiotics for the normal treatment of eosinophils. The treatment time is varied from 6 hours to 16 hours, however, eosinophils will undergo the cell death cycle so you need to be careful with that. I have never let my cells in the media for more than 24 hours.
In addition, we observe that the coculture of eosinophils with other types of cells can protect eosinophils survival, in that case, we just use the RPMI+1% antibiotics without serum.
Sorry for late reply!
Tu
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