非人灵长类多能干细胞体外培养和诱导分化的研究进展

doi:10.11843/j.issn.0366-6964.2024.10.004

Abstract

Abstract:

Stem cells have great value in disease occurrence, cell therapy, drug screening, and clinical applications. Since Shinya Yamanaka, a Japanese scientist, discovered induced pluripotent stem cells (iPSCs), the relevant research has become a research hotspot and up to the present. The study of mouse stem cells has opened up a path for the related research in other animals, but there are significant limitations in drug screening and disease treatment. Non-human primates have higher genetic homology with humans and are highly similar to humans in physiology, pathology, reproduction, pharmacology, nerves, and other aspects. They are ideal animal models for studying human diseases and developing drugs for human. Research on non-human primates is also a necessary step for a new drugs before entering clinical trials. The study of non-human primate stem cells is becoming important in the mechanism of diseases and drug development, while avoiding ethical issues. This article reviews the related research on non-human primate pluripotent stem cells including optimization of in vitro culture systems and methods of inducing differentiation, aiming to provide reference for further in-depth research on non-human primate stem cells.

Key words: non-human primate, pluripotent stem cells, in vitro culture, induced differentiation

CLC Number:

Q254

Kecheng SHEN, Jiaqiao ZHU, Zongping LIU. Research Progress on in vitro Culture and Induced Differentiation of Non-human Primate Pluripotent Stem Cells[J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(10): 4278-4289.

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种类 Type	名称 Name	成分 Ingredient	来源 Souece	特点 Peculiarity	应用 Application	参考文献 Reference
生物类基质 Biological matrix	明胶	明胶	胶原蛋白提取物	廉价, 但细胞存活率低，分化迅速	常用于定向分化	[12-13]
	Geltrex	主要为层黏连蛋白和Ⅳ胶原	鼠Engelbreth-Holm-Swarm肿瘤提取物	可维持NHP-PSCs 稳定传代不分化但有引入异源物质的风险	广泛适用于传代培养和定向分化
	Matrigel		小鼠肉瘤提取物			[14]
人工合成基质 Artificial synthetic matrix	PMEDSAH	氢氧化铵聚合物	人工合成	化学成分明确且能保持hESCs的增殖和未分化状态, 可以根据不同干细胞的特点定制	目前多用于各种人源干细胞的培养，很少用于NHP-PSCs的培养	[15-17]
	APMAAm 水凝胶	氨基丙基甲基丙烯酰胺	人工合成

细胞系 Cell line	目标细胞 Goal	诱导方法 Method	诱导条件 Conditions	应用效果 Application effect	参考文献 Reference
ESC系	血管母细胞	拟胚体分化法	BMP4、FLT3L、SCF、血小板生成素、FGF、VEGF和KSR	可将拟胚体分化为血管母细胞，后将其接种到体外培养的损伤股动脉中，可进一步分化成血管内皮细胞	[49]
iPSCs系	心肌细胞	分步诱导法	Matrigel基质胶、激活素A和BMP4	诱导分化而来的心肌细胞经同种移植后可以改善心脏收缩功能，但移植受体会出现心动过速	[50]
ESC系和iPSCs系	造血细胞	拟胚体分化法和基质细胞共培养法	FBS、PSG、ITS、MTG、抗坏血酸磷酸酯、VEGF，以及SCF和FLT3L	可以将PSCs诱导分化为造血祖细胞，再用细胞因子或T细胞受体诱导成各种类型的T细胞	[51]

细胞系 Cell line	目标细胞 Goal	诱导方法 Method	诱导条件 Conditions	应用效果 Application effect	参考文献 Reference
ESC系ESC	神经祖细胞	拟胚体分化法	ITS、纤连蛋白、黏连蛋白、bFGF和N-2	90%为表达巢蛋白(nestin)的神经祖细胞，但胰腺前体细胞和某些成熟组织也表达nestin，需要其他标志物如SOX2、occludin等共同确定	[52-58]
iPSCs系iPSCs	多巴胺分泌神经细胞	基质细胞共培养法	KSR, 丙酮酸盐, 非必需氨基酸, 2-巯基乙醇	35%的细胞被成功诱导分化，但诱导效率仅仅是诱导小鼠ESCs的一半	[59-60]
ESC系ESC	端脑神经细胞	拟胚体分化法	Wnt抑制剂, Nodal抑制剂、Wnt3a、Shh	可定向诱导为大脑皮质或大脑基底核	[37, 61]

细胞系 Cell line	目标细胞 Goal	诱导方法 Method	诱导条件 Conditions	应用效果 Application effect	参考文献 Reference
ESC系ESC	肝细胞样细胞	拟胚体分化法	aFGF、HGF、Y-27632、FGF-4、BMP2、激活素A、ITS、肿瘤抑制因子、人胰岛素和地塞米松等	诱导细胞呈现肝细胞特异性形态，而且不同程度的表达多种肝细胞标志分子	[96-100]
ESC系和iPSCs系ESC and iPSCs	胰岛素分泌细胞	多步诱导法	激活素A和WNT3A，RA、KGF、EGF、SB431542、HGF、IFG1、exendin4和烟酰胺等	细胞表达胰岛细胞的标志分子，移植到糖尿病模型小鼠后能使50%的小鼠恢复血糖	[101]
iPSCs系iPSCs	肌源性祖细胞	多步诱导法	过表达PAX7、SB-431542、LDN-193189、dox、LY-374973、forskolin等	细胞最终分化为肌管；将肌源性祖细胞分别移植到小鼠和猴体内能促进肌肉的再生	[102]
ESC系ESC	类囊胚	多步诱导法	bFGF、激活素A、CHIR99021、PD0325901、A83-01、IWR1、TSA、DZNep等	成功实现了食蟹猴类囊胚的体外构建；经同种移植后，这一类囊胚可以着床并引发妊娠反应	[103]

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Research Progress on in vitro Culture and Induced Differentiation of Non-human Primate Pluripotent Stem Cells

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