烏骨雞誘導多能性幹細胞株體外分化能力之探討

Evaluation of in vitro and in vivo differentiation capability of induced pluripotent stem cell lines from the black silkie chicken

劉振發(Jenn-Fa Liou) ； 陳裕信(Yu-Hisn Chen) ； 蕭振文(Jen-Wen Shiau) ； 薛佑玲(Yow-Ling Shiue) ； 陳立人(Lih-Ren Chen)

《畜產研究》 52卷4期 (2019/12) Pp. 215-226

摘要

本研究目的為探討烏骨雞誘導多能性幹細胞（black silkie chicken induced pluripotent stem cells, BSciPSCs）之體外分化特性和嵌合體形成能力，期供後續相關研究使用。供試之BSciPSCs在體外培養已超過35繼代（約300天），經分化多能性專一性抗體Oct-4、AP及PAS染色後可呈現陽性反應，且利用胚層細胞譜系特異性抗體進行免疫染色方式檢驗BSciPSCs衍生的類胚體（EB）的結果顯示，BSciPSCs具有分化形成三胚層的細胞譜系和神經細胞的能力。將BSciPSCs移植到孵化3.5天（X-stage）的來亨雞胚中，檢視孵化第12天胚胎的體色分佈，結果有11%胚胎體色有黑色分佈，證實為嵌合體。綜合上述結果，在本研究中建立的BSciPSCs是具多能性，且有嵌合體形成潛力，冀望此細胞可應用於生物醫學領域研究。

關鍵字

黑絨烏骨雞 ； 誘導多能性幹細胞 ； 嵌合體

並列摘要

The purpose of this study was to evaluate the in vitro and in vivo differentiation capability of the induced pluripotent stem cells (iPSCs) derived from the black silkie chicken (BSciPSCs). The BSciPSCs have been maintained in vitro for more than 35 passages (about 300 days). These BSciPSCs continuously expressed pluripotent markers of stem cells including Oct- 4, AP, and PAS antigens. The in vitro differentiation ability of BSciPSCs was determined by EB formation and spontanely differentiation induction, and the resultant lineages were verified by immunostaining with antibodies specific against cells from 3 primitive germ layers and neuron cells. Moreover, the in vivo differentiation capability of BSciPSCs was further demonstrated by formation of multiple lineaged tetramos in NOD-SCID mice and resulted 11% chimeric chicken embryos after transplantiation of BSciPSCs into Leghorn chicken embryos. In conclusion, the BSciPSC lines established in this study can be maintained in vitro continuously without losing their in vitro and in vivo pluripotency. The success of BSciPSCs estabilishment would provide an alternative modles for biomedicine study.

並列關鍵字

Black silkie chicken ； Induced pluripotent stem cells ； Chimera