Possibility to Establish Chicken Stem Cell from Non-germline Tissue; Detection of Colony-forming Cells after Chicken Fibroblast Culture and Subsequent Stem Cell Characterization

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This study was conducted to evaluate whether culture of chicken fibroblasts could induce cell aggregation leading to embryonic stem cell (ESC)-like, colony-formation. Gradual increase in expression of leukemia inhibitory factor (LIF) gene were detected as retrieval age of chicken embryonic fibroblasts (CEFs) was increased from 5- to 8-day-old, which accompanied increase in cell proliferation after culture in LIF-free medium. Colony-like cell aggregation was detected after culture of the 8-day-old fibroblasts, while the exposure to 0.5, 1 or 2 mM glutathione did not increase the cell aggregation. The cell aggregates were positive for periodic acid Schiff solution, and the antibodies of anti-stage specific embryonic antigen (SSEA)-1 and anti-SSEA-3. However, they were negative for alkaline phosphatase and anti- SSEA-4 antibody. Expression of chicken Vasa homolog did not detect in the aggregated CEFs. Continuous culture of the aggregates in LIF- and CEF monolayer-free condition resulted the formation of embryoid bodies possessing the cells being positive for three germ layer makers (Nestin, S-100, smooth muscle actin, desmin, alpha-fetoprotein and Troma-1). In conclusion, our results demonstrate cell aggregates having ESC-like activity can be derived from the culture of chicken fibroblasts on CEF monolayer, which strongly supports the possibility on deriving chicken stem cells from non-germline tissue.
日本家禽学会の論文