家禽胚胎干细胞的研究与应用进展

doi:10.19543/j.cnki.1673-4556.20260803.002

Abstract

Abstract:

Research on poultry embryonic stem cells, as an important interdisciplinary field bridging developmental biology and agricultural biotechnology, has achieved remarkable progress in recent years. This review systematically summarizes the fundamental characteristics, key technological breakthroughs, current challenges, and potential applications of poultry embryonic stem cells, with a focus on recent advances in the molecular mechanisms underlying pluripotency maintenance, optimization strategies for culture systems, application of gene-editing technologies, and generation of interspecies chimeras. At the molecular level, the core pluripotency regulatory network of poultry embryonic stem cells is composed of POU5F3, SOX3, and NANOG, which differs significantly from that of mammals. In particular, novel culture systems such as OT/2i and OT/3i/chLIF are highlighted. By precisely inhibiting differentiation-promoting signaling pathways including Wnt, PKC, and Activin/TGF-β, and supplementing with chicken leukemia inhibitory factor （chLIF）, these systems provide long-term, stable culture support for embryonic stem cells derived from multiple avian species, including chicken, quail, duck, and goose. On this basis, the CRISPR-Cas9 gene-editing technology has been successfully applied to poultry embryonic stem cells, enabling efficient gene knock-in and site-specific modification. Furthermore, the successful generation of interspecies chimeras and the confirmation of germ cell differentiation capacity have further validated the developmental pluripotency of these cells. Finally, based on a comprehensive synthesis of existing research findings, this review identifies major challenges in the field, such as low germline transmission efficiency and insufficient long-term genetic stability, and offers a forward-looking perspective on future developments.

Key words: Embryonic stem cells, Poultry, In vitro culture, Pluripotency, Interspecies chimera, Gene editing

CLC Number:

Youchen Liang, Jianxiao Zhao, Qimeng Zhang, Zhi Cao, Qian Xue, Guohui Li, Hongyan Sun, Yinjie Niu, Qisheng Zuo, Wei Han, Bichun Li, Kai Jin. Research and application progress of poultry embryonic stem cells[J].Chinese Livestock and Poultry Breeding, 2026, 22(9): 26-36.

Figures/Tables 8

Table 1

Fig.1

Table 2

Common molecular markers of avian ES cells"

类别 Category	分子标记物 Molecular marke	表达特征 Expression pattern	功能意义 Functional significance	检测技术 Detection method	参考文献Reference
核心转录因子 Core transcription factors	POU5F3 （PouV）	细胞核高表达	维持多能性核心调控因子，替代哺乳动物OCT4功能	免疫荧光、Western Blot	[8]
	NANOG	细胞核高表达	维持自我更新，抑制分化	免疫荧光、RT-qPCR	[12]
	SOX3	细胞核高表达	替代哺乳动物SOX2功能，参与多能性调控	免疫荧光、RNA-seq	[4]
	SALL4	细胞核高表达	与POU5F3协同调控多能性网络	免疫荧光、RT-qPCR	[8]
	KLF2	细胞核高表达	维持多能性，抑制分化；支持自我更新	RT-qPCR、免疫荧光、RNA-seq	[9-11]
	KLF5	细胞核高表达	维持多能性的关键调控因子，抑制分化且在禽类中取代哺乳动物KLF4的核心作用，促进多能性网络稳定	RT-qPCR、Western Blot、免疫荧光	[9-11]
	KLF6	细胞核高表达	与KLF5协同，调控多能性基因并参与抑制分化路径	RNA-seq、RT-qPCR、免疫荧光	[9-11]
	KLF4	细胞核低表达	在chESCs中非主导，但部分参与核心网络。整体在禽类中被KLF2/5/6部分取代	RT-qPCR、RNA-seq	[9-11]
表面抗原 Surface antigens	SSEA-1	细胞膜阳性	未分化状态标志，表达呈异质性	流式细胞术、免疫荧光	[7]
	EMA-1	细胞膜阳性	禽类特异性表面抗原	流式细胞术、免疫荧光	[13]
	Integrin α6β1	细胞膜高表达	介导细胞与基质粘附，维持干细胞微环境	流式细胞术	[14]
功能蛋白 Functional proteins	碱性磷酸酶	高活性	未分化状态经典标志	组织化学染色	[10]
	LIN28A/B	细胞质高表达	调控microRNA，维持多能性	Western Blot、RT-qPCR	[4]
	TERT	高表达	维持端粒长度，支持长期增殖	RT-qPCR	[10]
生殖细胞标记 Germ cell markers	DAZL	分化后表达	生殖细胞分化标志	免疫荧光、RT-qPCR	[8]
生殖细胞标记 Germ cell markers	CVH	分化后表达	禽类VASA同源物，生殖细胞特异	原位杂交、RT-qPCR	[15]

Table 2

Table 3

Fig. 2

Fig. 3

Table 4

Fig. 4

References 36

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类胚体标记基因Embryoid body marker genes	关键基因Critical genes	表达水平Expression level	参考文献References
外胚层 Ectoderm	NeuroG1	高	[4]
	Sox1	高	[4,8]
	Nestin	高	[4,18]
中胚层 Mesoderm	Gsc	高	[4]
	T	高	[4]
	Pdgfa0	高	[4]
内胚层 Endoderm	Gata6	高	[4]
	Pdx1	高	[4]
	Foxa2	高	[4]
多能性基因 Pluripotency genes	Nanog	低	[4,8,11-12]
	Pou5f3	低	[4]
	Sall4	低	[4,8]

培养体系Culture system	名称 Name	功能和意义 Function and significance
OT/2i	卵转铁蛋白	卵转铁蛋白是卵黄中维持cESCs自我更新的最关键成分
	IWR-1	一种Wnt/nt/1最关键成分信号通路抑制剂，适度的Wnt信号抑制对维持禽类多能态至关重要
	Go6983	一种PKC信号通路抑制剂，与IWR-1协同作用，压制诱导分化的信号
OT/3i	SB431542	通过抑制Activin/TGF-i受体ALK4/5/7来阻止这一分化路径
OT/3i/chLIF	chLIF	通过激活JAK/STAT3通路有效抑制cESCs的分化

Research and application progress of poultry embryonic stem cells

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