Research Progress in the Application and Establishment of Bovine Induced Pluripotent Stem Cells

doi:10.13560/j.cnki.biotech.bull.1985.2022-1371

Abstract

Abstract:

Induce pluripotent stem cells（iPSCs）refer to a class of stem cell lines that reprogram terminally differentiated somatic cells into infinite proliferation and regeneration and have multiple cell types differentiated into three germ layers by introducing specific transcription factors. At present, many important achievements have been made in the research of iPSCs in humans and mice, but no breakthrough has been made in the research of iPSCs in other animals, such as cattle and other economic ungulates. How to efficiently and safely introduce foreign transcription factors into somatic cells through reprogramming vectors and continuously express them is the main bottleneck in the production of bovine induced pluripotent stem cells. This article reviews the selection of reprogramming system, induction factors and small molecule compounds in the establishment of bovine iPSCs, aiming to provide reference for further improvement of biPSCs and the establishment of bovine embryonic stem cell lines.

Key words: bovine induced pluripotent stem cells, somatic cell reprogramming, cultivation system, small molecular compounds

ZENG Hong, ZENG Rui-lin, FU Wei, JI Wen-hui, LAN Dao-liang. Research Progress in the Application and Establishment of Bovine Induced Pluripotent Stem Cells[J]. Biotechnology Bulletin, 2023, 39(5): 130-141.

Figures/Tables 5

References 86

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非整合型方法 Approach of non-integrating	优点 Pros	缺点 Cons
腺病毒	基因组整合频率低，载体容量大	重编程效率低，具有免疫原性，对某些细胞需多次感染
仙台病毒	无基因组整合，载体容量大，转染效率高，稳定转染，重编程效率高	多个转录因子需多个病毒携带，具有免疫原性，需传代10次以上才可完全清除病毒基因组，只能加工RNA
质粒转染	基因组整合频率低，操作简单，免疫原性低，无病毒组分	不能自我复制，重编程效率极低，需要多次转染
转座子	不涉及病毒，可精确切除，只需一次转染，载体容量大，免疫原性低	可能导致突变和染色体重排，重编程效率低，需要用转座酶进行切除
微环DNA载体	基因组整合频率低，操作简单，转染效率高，免疫原性极低	不能自我复制，需多次转染，重编程效率极低
小分子化合物	具有瞬时控制能力，易操作，能促进iPSCs的生成	对毒性效率存在争议，确定或非特异的效果具有不确定性或非特异性

非整合型方法 Approach of non-integrating	优点 Pros	缺点 Cons
腺病毒	基因组整合频率低，载体容量大	重编程效率低，具有免疫原性，对某些细胞需多次感染
仙台病毒	无基因组整合，载体容量大，转染效率高，稳定转染，重编程效率高	多个转录因子需多个病毒携带，具有免疫原性，需传代10次以上才可完全清除病毒基因组，只能加工RNA
质粒转染	基因组整合频率低，操作简单，免疫原性低，无病毒组分	不能自我复制，重编程效率极低，需要多次转染
转座子	不涉及病毒，可精确切除，只需一次转染，载体容量大，免疫原性低	可能导致突变和染色体重排，重编程效率低，需要用转座酶进行切除
微环DNA载体	基因组整合频率低，操作简单，转染效率高，免疫原性极低	不能自我复制，需多次转染，重编程效率极低
小分子化合物	具有瞬时控制能力，易操作，能促进iPSCs的生成	对毒性效率存在争议，确定或非特异的效果具有不确定性或非特异性

组别 Group	Naïve状态 Naïve state	Primed状态 Primed state
培养条件	LIF/血清或2i	Activin A/bFGF或CHIR/IWR1
FGF2依赖性	不依赖	依赖
LIF依赖性	依赖	不依赖
BMP4依赖性	依赖	不依赖
Wnt激活	自我更新	分化
克隆形态	隆起	扁平
XX失活情况	XaXa	XaXi
Nanog表达	高	低
Fgf5表达	低	高
OCT4增强子	远端	近端
H3K24me3表达	低	高
嵌合小鼠胚胎	能	不能

组别 Group	Naïve状态 Naïve state	Primed状态 Primed state
培养条件	LIF/血清或2i	Activin A/bFGF或CHIR/IWR1
FGF2依赖性	不依赖	依赖
LIF依赖性	依赖	不依赖
BMP4依赖性	依赖	不依赖
Wnt激活	自我更新	分化
克隆形态	隆起	扁平
XX失活情况	XaXa	XaXi
Nanog表达	高	低
Fgf5表达	低	高
OCT4增强子	远端	近端
H3K24me3表达	低	高
嵌合小鼠胚胎	能	不能

培养基 Medium	主要添加成分 Main component	参考文献 Reference
CTRF	TeSR1培养基（无FGF2和TGFb）、FGF2和IWR1	[4]
LCDM	hLIF、CHIR99021、（S）-（+）-dimethindene maleate、minocycline hydrochloride	[60]
TiF培养基	mTeSR plus培养基、PD0325901、CHIR-99021、hLIF、IWR1、iDOT1L、Forskolin	[44]
bEPSC培养基	mTeSR1培养基、CHIR99021、WH-4-023、XAV939、IWR-1、维生素C、LIF、Activin A	[19]