Feeder Removal MicroBeads have been developed for the efficient depletion of mouse feeder cells from co-cultures with human or mouse ES and iPS cell lines and can also be used with other, feeder-dependent cell lines. Depletion of feeder cells, e.g. prior to differentiation or gene expression profiling studies, helps to obtain reproducible results.
The most prevalent cultivation method for maintenance of mouse and human embryonic stem (ES) cells and induced pluripotent stem (iPS) cells is coculturing on mouse embryonic fibroblast feeder cell layers. Different feeder cell types are employed, among these primary embryonic fibroblasts (mEF) and mEF-derived cell lines (e.g. NIH3T3). Feeder cells are also used in maintenance cultures of primary keratinocytes.
Depletion of feeder cells improves the reproducibility of many downstream application, including
- Gene expression-profiling
- Blastocyst injection of ES cells after gene targeting
- Differentiation protocols for ES and iPS cells: Removal of feeder cells can improve the responsiveness to developmental cues.
LS or autoMACS® Columns.
Mouse embryonic fibroblasts (mEF) were depleted from a co-culture with mouse embryonic stem (mES) cells using Feeder Removal MicroBeads, an LS Column, and a MidiMACS™ Separator. Flow cytometric analysis with CD15 (SSEA-1)-APC and Anti-Feeder-PE on the MACSQuant® Analyzer indicates that a starting population of 10% mEF was completely removed. The purity of the enriched mES cells was about 99.9%.