Culture conditions for bovine embryonic stem cell-like cells isolated from blastocysts after external fertilization
Although isolation and characterization of embryonic stem cells have been successful in cattle, maintenance of bovine embryonic stem cells in culture remains difficult. In this study, we compared different methods of cell passaging, feeder cell layers and medium conditions for bovine embryonic stem cell-like cells. We found that a murine embryonic fibroblast feeder layer is more suitable for embryonic stem cell-like cells than bovine embryonic fibroblasts. When murine embryonic fibroblasts were used, a mechanical method of passaging led to better cell growth than passaging by trypsin digestion. We also found that exogenous supplementation with leukemia inhibitory factor maintained the embryonic stem cell-like cells in an undifferentiated state, whereas addition of stem cell factor resulted in their differentiation. Our findings provide an experimental basis for the establishment of an effective culture system for bovine embryonic stem cells.
KeywordsBovine embryonic stem cell-like cells Feeder layer Passage SCF LIF
This work was supported by grants from the National High Technology Research and Development Program of China (“863” Program) (2008AA101005). We gratefully thank Dr. Cang Ming (Inner Mongolia University) and Dr. Guo Xudong (Inner Mongolia University) for providing the directions.
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