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中國畜牧學會會誌
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中文摘要
本試驗旨本研究目的為建立雞誘導多能性幹細胞（chicken induced pluripotent stem cells， ciPSC），並探討雞誘導多能性幹細胞之分化多能性、胚體形成效率與分化潛力等特性，期 供後續相關研究使用。試驗結果顯示，利用慢病毒（lentivirus）將轉錄因子基因 LIN28, NANOG, SOX2, OCT3/4, KLF4 和 c-MYC 轉染雞胚纖維母細胞（chicken embryo fibroblasts；cEFs）6-7 天後， cEFs 之細胞形態逐漸由梭狀轉為圓形之上皮細胞形態，持 續培養 21 天後形成類似幹細胞群落形態。ciPSC 體外培養已超過 35 代（300 天），經 分化多能性專一性抗體 Oct-4、AP、與 PAS 染色後可呈現陽性反應；在形態學上亦呈現 具明顯細胞邊界、高核質比及大細胞核仁等幹細胞特徵。使用懸浮小滴培養技術培養，具 有高效率（92.6 ± 2.2%）的類胚體（embryoid body）形成率；顯示所建立之雞誘導多能性 幹細胞具有分化多能性之潛能。我們已成功建立 ciPSC，冀望此細胞可應用於生物醫學和 人類疾病領域研究。
英文摘要
The purpose of this study is to establish chicken induced pluripotent stem cells (ciPSC) line and to study their celluar characters including pluripotency, embryoid body formation efficiency, and in vitro differentiation capability. The results showed that the morphology of chicken embryo fibroblasts (cEF) transformed into colony type from spindle type 6-7 days after being infected with lentivirus, which was constructed with reprogramming transcription factors of LIN28, NANOG, SOX2, OCT3/4, KLF4 and c-Myc. The transformed cells have been maintained in vitro for more than 35 passages (300 days). These ciPSCs continuously expressed pluripotent markers of stem cells including Oct-4, AP, and PAS antigens. Morphologically, ciPSC colonies were highly refractive, and at the single-cell level they showed clear cell boundary, high nuclear-to-cytoplasm ratio, and prominent nucleoli. The efficiency of embryoid body formation (92.6 ± 2.2 %) was excellent induced by handing-drop culture. These results demonstrated that the ciPSC line established in this study were pluripotent.