RBL-2H3 line - (Jul/20/2005 )
Hi, all:
Am wondering if any one is familiar with culturing this line...
Our lab recently acquired this line and was told that we should use DMEM (high glucose) with 10% FBS plus pen/strep to culture it. Initially, the cells grew ok and we've successfully frozen some for the storage purpose.
However, after about a month or so, we started noticing that the cells are gradually becoming slow-growers (in that, with a 25% confluency, it took more than 2 weeks for them to become confluent). We haven't seen any contamination except there seemed to be more cells floating in the flask (this cell line is the adherent type). We went up to ATCC's website and found out the media suggested by ATCC is as follows:
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Minimum essential medium (Eagle) with 2 mM L-glutamine and Earle's BSS adjusted to contain 1.5 g/L sodium bicarbonate, 0.1 mM non-essential amino acids, and 1.0 mM sodium pyruvate, 85%; heat-inactivated fetal bovine serum, 15%
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Do any expert with experience on this line kindly give me some advice as whether the DMEM media we used is ok for this line. And, if there's anything we could check to see if something might have gone wrong during our culturing it.. Many thanks..
Hi Lab Newbie,
I work with the RBL-2H3 cell line too. I am facing problems in the revival of the cryo-preserved stock. i use 5% DMSO in 95% RPMI-1640 medium for the cryo medium.
I culture the cells in RPMI-1640 medium too.
Do you have any suggestions as to what might be the problem and could you givee somew tips please for the revival of teh cell line? thanks.
in
Am wondering if any one is familiar with culturing this line...
Our lab recently acquired this line and was told that we should use DMEM (high glucose) with 10% FBS plus pen/strep to culture it. Initially, the cells grew ok and we've successfully frozen some for the storage purpose.
However, after about a month or so, we started noticing that the cells are gradually becoming slow-growers (in that, with a 25% confluency, it took more than 2 weeks for them to become confluent). We haven't seen any contamination except there seemed to be more cells floating in the flask (this cell line is the adherent type). We went up to ATCC's website and found out the media suggested by ATCC is as follows:
---
Minimum essential medium (Eagle) with 2 mM L-glutamine and Earle's BSS adjusted to contain 1.5 g/L sodium bicarbonate, 0.1 mM non-essential amino acids, and 1.0 mM sodium pyruvate, 85%; heat-inactivated fetal bovine serum, 15%
---
Do any expert with experience on this line kindly give me some advice as whether the DMEM media we used is ok for this line. And, if there's anything we could check to see if something might have gone wrong during our culturing it.. Many thanks..