We just got MUTZ-3 cell line in and wanted to check their cell surface markers -comparing undifferentiated to differentiated and stimulated etc. However we seem to be getting high levels (Aprox. 60%) of CD86 and HLA-DR on the undifferentiated cells. I would expect some expression but no where near that high and was wondering what sort of levels others normally see? Could the way we maintain the cells be in any way responsible for this?
Hi Amy,
Cell lines with high passage number gets differentiated over time and start behaving abnormal. First check if that could be one reason. Check ur culturing conditions that are needed for maintaining MUTZ-3 cell lines. Also have a go thorough below mentioned articles which may give an extra bit of information than you needed.
http://www.jleukbio.org/content/84/6/1364.full.pdf+html
http://www.bloodjournal.org/content/bloodjournal/100/2/701.full.pdf?sso-checked=true
Hi Amy, those are perfectly normal levels of expression on this cell line. Once you start differentiation induction percentages and levels (at least of CD86) will go up in conjunction with other co-stim molecules and DC markers like CD1a.
Hi, thanks for the information Tanja. I guess the reason I was surprised at the levels being that high was because the datasheet on the companies website (creative bioarray) for HLA-DR on undifferentiated MUTZ-3 says that the levels are 35%...and we have seen several other papers saying that CD86 is really low on both undifferentiated and immature MUTZ-3 cells. We really need to know if the levels we are seeing are normal or not as our experiment relies on getting the cells to be immature but no further in their differentiation. Do you have any links to papers that show them with these higher levels of CD86/HLA-DR?
Hi Amy:
Santegoets et al. J Leukoc Biol. 2006 Dec;80(6):1337-44. Epub 2006 Sep 7.
PMID: 16959899 [PubMed - indexed for MEDLINE] Free Article
http://www.ncbi.nlm.nih.gov/pubmed/16959899
This paper provides a very useful Table with marker expression levels on MUTZ-3 precursors (CD34 vs double neg vs CD14 positive) .
Other useful papers may be
Santegoets et al J Immunol 2008 Apr 1;180(7):4540-9.
PMID: 18354176 [PubMed - indexed for MEDLINE]
Santegoets et al J Leukoc Biol. 2008 Dec;84(6):1364-73. doi: 10.1189/jlb.0208092. Epub 2008 Jul 29. Review.
PMID: 18664532 [PubMed - indexed for MEDLINE]
good luck!
Tanja
We established and characterized this cell line. CB copied our data and applied to cells of uncertain provenance since we fused to supply them with original cells. hence, I wonder which cells they are actually supplying. Perform STR profiling and compare them with reference profiles at DSMZ where I work.