Joanne Yetz-Aldape, I have looked at adipocytes, particularly 3T3L1's, using flow, and they are relatively easy to keep in a single cell suspension. Since we also did'nt have surface markers for differentiation, we measured lipid accumulation using Nile Red. A time study revealed a full peak shift by day 7. This method may be useful for evaluating changes in cell differentiation patterns. I would like to know also what kind of surface markers can be used for adipocytes. By the way, human adipocytes were not as easy to work with because they are more stubborn about de-attachnig and tend to break apart when fully mature. Here is a simplified protocol: trypsinize cells, gather in 750ul total volume,12x75 tubes do not wash add 250ul 2% paraformaldehyde (final = 0.5%) incubate 10 min add 100ul Nile Red (final = 1ug/ml) run FACS using FL2 585nm filter Good luck, Cissy Geigerman On Fri, 27 Oct 2000 15:38:49 -0400 Joanne Yetz <jyetz@genetica.cc> wrote: > > Hello Flow-ers, > > I was wondering if anyone had any experience looking at adipocytes, > specifically the 3T3-L1 cell line, on a flow cytometer. Are these cells > particularly difficult to prepare and maintain in a single cell suspension > for cell sorting? Are there any cell surface markers which are useful for > phenotyping these cells? Are there any markers which appear or disappear as > the cell differentiates? I am looking for some marker(s) that I could > monitor and use as an assay for differentiation of the 3T3-L1 cell line. > Any information would be greatly appreciated. Thanks. > > Joanne Yetz-Aldape > Genetica, Inc. > One Kendall Square Building 600 > Cambridge, MA 02139 > jyetz@genetica.cc > (617) 621-1222 x245 > ---------------------- Cissy Geigerman Dept.of Nutrition and Food Service Systems Univ. of North Carolina at Greensboro (336)334-5313
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