BM cells were collected from femurs and tibiae in X-VIVO 20 media (Cambrex, CA) for analysis and sorting and stained with the indicated combination of antibody.
After staining, the cells were centrifuged and resuspended in HBSS (Invitrogen) containing 1 µg/ml propidium iodide and maintained at 4°C for flow cytometry analysis and sorting.
The cells were then placed in a MoFlo High-Performance Cell Sorter (Dako Cytomation, Glostrup, Denmark) using Summit 4.0 Software (Dako Cytomation) for analysis and sorting.
On specified days of co-culture, cells were recovered from the culture dishes for analysis and sorting on a flow cytometer.
The cells at the interphase were collected and, after washing with HBSS, stained with monoclonal antibodies for flow cytometry analysis and sorting.
We thank the Alvin J. Siteman Cancer Center at Washington University School of Medicine and Barnes-Jewish Hospital in St . Louis MO, for the use of the High Speed Cell Sorter Core, which provided flow cytometry analysis and sorting services and for the use of the Multiplexed Gene Analysis Core, which provided Affymetrix GeneChip microarray services.
OVB, DBS, AAK, and GVS worked with blood samples and performed flow cytomerty analysis and sorting.
The cell pellets were resuspended in FACS buffer for further analysis and sorting.
Analysis of control E14 ES cells were used as a negative control for flow analysis and sorting.
The resulting double-emulsion droplets are suitable for quantitative analysis and sorting in a commercial flow cytometer.
