提高猪体细胞克隆胚胎发育率的研究进展

doi:10.13304/j.nykjdb.2024.0209

中国农业科技导报 ›› 2024, Vol. 26 ›› Issue (12): 201-209.DOI: 10.13304/j.nykjdb.2024.0209

• 方法与技术创新 • 上一篇

提高猪体细胞克隆胚胎发育率的研究进展

张才用¹(), 陈指龙², 谢浩², 彭翠婷², 晏超², 赵玉兰², 齐霖², 周磊¹, 唐中林²^,³()

^1.广西大学动物科学技术学院，南宁 530000
^2.中国农业科学院（深圳）农业基因组研究所，岭南现代农业科学与技术广东省实验室深圳分中心，农业农村部畜禽生物组学重点实验室，广东深圳 518000
^3.佛山鲲鹏现代农业研究院，广东佛山 528225

收稿日期:2024-03-19 接受日期:2024-04-18 出版日期:2024-12-15 发布日期:2024-12-17
通讯作者: 唐中林
作者简介:张才用 E-mail：1797832600@qq.com；
基金资助:
深圳市创新创业计划-科技重大专项项目(KJZD20230923115003006);国家自然科学基金青年项目(32302747);广东省基础与应用基础研究基金委员会区域联合基金-青年基金项目(2021A1515110046);巴马县人才科技计划项目(巴人科20210026)

Progress of the Improvement of Development of Porcine Somatic Cell Nuclear Transfer Embryos

Caiyong ZHANG¹(), Zhilong CHEN², Hao XIE², Cuiting PENG², Chao YAN², Yulan ZHAO², Lin QI², Lei ZHOU¹, Zhonglin TANG²^,³()

^1.College of Animal Science and Technology，Guangxi University，Nanning 530000，China
^2.Shenzhen Branch Center of Guangdong Laboratory of Lingnan Modern Agricultural Science and Technology，Key Laboratory of Livestock and Poultry Biohistology，Ministry of Agriculture and Rural Development，Agricultural Genomics Institute at Shenzhen，Chinese Academy of Agricultural Sciences，Guangdong Shenzhen 518000，China
^3.Kunpeng Institute of Modern Agriculture at Foshan，Guangdong Foshan 528226，China

Received:2024-03-19 Accepted:2024-04-18 Online:2024-12-15 Published:2024-12-17
Contact: Zhonglin TANG

摘要/Abstract

摘要：

体细胞克隆技术可使终末分化的体细胞获得完整全能性，进而生产出完整个体。猪体细胞克隆技术在种质资源的保护与利用及制备基因编辑猪等方面具有广泛的应用前景。然而，猪体细胞克隆的胚胎发育率和移植存活率限制着该技术在实际生产中的推广应用。随着微量细胞测序技术的发展，克隆胚胎发育异常机制不断被揭示，诸多研究人员尝试通过改良核移植操作程序或纠正克隆胚胎的异常重编程来提高克隆成功率，取得了一定效果。综述了近年来围绕卵母细胞、供体细胞和克隆胚胎3个方面来提高猪体细胞克隆胚胎发育率的相关研究，以期为提高猪体细胞克隆效率提供参考，有助于推动体细胞克隆技术在猪生产中的应用。

关键词: 猪, 体细胞, 核移植, 胚胎, 发育

Abstract:

Somatic cell cloning technology can driver the somatic cells of terminally differentiated to obtain totipotency， it has the potential to develop into the living individual. The technology of porcine somatic cell cloning has a wide range of applications for the conservation and utilization of germplasm resources and the production of gene-edited pigs. However， the low rate of cloned embryo development and transfer survival limit the popularization of this technology in practical production. With the development of sequencing technology for microcell， the mechanism of abnormal development of cloned embryos has been revealed， and many researchers have tried to improve the success rate by modifying the nuclear transfer procedure or correcting the abnormal reprogramming of cloned embryos， which has achieved remarkable results. The research progress on the improvement of cloning efficiency through oocytes， donor cells and cloned embryos in recent years were summarize， which aimed at providing reference to improve the efficiency of porcine somatic cell cloning and would help to promote the application of somatic cell cloning technology in swine production.

Key words: pig, somatic cell, nuclear transfer, embryo, development

中图分类号:

S828

张才用, 陈指龙, 谢浩, 彭翠婷, 晏超, 赵玉兰, 齐霖, 周磊, 唐中林. 提高猪体细胞克隆胚胎发育率的研究进展[J]. 中国农业科技导报, 2024, 26(12): 201-209.

Caiyong ZHANG, Zhilong CHEN, Hao XIE, Cuiting PENG, Chao YAN, Yulan ZHAO, Lin QI, Lei ZHOU, Zhonglin TANG. Progress of the Improvement of Development of Porcine Somatic Cell Nuclear Transfer Embryos[J]. Journal of Agricultural Science and Technology, 2024, 26(12): 201-209.

图/表 3

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化合物 Compound	用量 Dosage	时间 Time	作用机制 Mechanism	囊胚率（处理vs对照） Blastocyst （treated vs control）/%
喹诺他^［56］ Quisinostat	10 nmol·L^-1	24 h	调节POU5f1蛋白、BAX和BCL2基因的表达和表观遗传修饰，促进供体细胞核重编程 Regulate the expression of POU5f1 protein， BAX， BCL2 genes and epigenetic modification， promote nuclear reprogramming in donor cells	（19.0±1.6） vs （10.2±0.9）
Scriptaid^［57］	100 nmol·L^-1	24 h	提高克隆胚胎的组蛋白去乙酰化水平，促进核的重编程 Increase histone deacetylation in cloned embryos promotes nuclear reprogramming	30.4 vs 17.5
MGCD0103^［58］	0.2 µmol·L^-1	24 h	提高假原核期胚胎H3K9ac和H4K12ac的修饰水平，增强SCNT胚胎的核重编程能力 Increase the levels of H3K9ac and H4K12ac in pseudoprokaryotic stage and enhance nuclear reprogramming in SCNT embryos	25.5 vs 10.7
Oxamflatin^［59］	1 µmol·L^-1	15 h	提高多能基因POU5F在囊胚阶段的表达和2-细胞和4-细胞期的组蛋白H3K9和H4K5乙酰化修饰水平，降低重构胚胎原核期去乙酰化酶活性和2-细胞期的DNA甲基化水平Increase expression of the pluripotency gene POU5F at the blastocyst stage and histone H3K9 and H4K5 acetylation modification levels at the 2-cell and 4-cell stages， and decrease deacetylase activity at the prokaryotic stage of remodeled embryos and DNA methylation levels at the 2-cell stage	23.6 vs 11.7

化合物 Compound	用量 Dosage	时间 Time	作用机制 Mechanism	囊胚率（处理vs对照） Blastocyst （treated vs control）/%
喹诺他^［56］ Quisinostat	10 nmol·L^-1	24 h	调节POU5f1蛋白、BAX和BCL2基因的表达和表观遗传修饰，促进供体细胞核重编程 Regulate the expression of POU5f1 protein， BAX， BCL2 genes and epigenetic modification， promote nuclear reprogramming in donor cells	（19.0±1.6） vs （10.2±0.9）
Scriptaid^［57］	100 nmol·L^-1	24 h	提高克隆胚胎的组蛋白去乙酰化水平，促进核的重编程 Increase histone deacetylation in cloned embryos promotes nuclear reprogramming	30.4 vs 17.5
MGCD0103^［58］	0.2 µmol·L^-1	24 h	提高假原核期胚胎H3K9ac和H4K12ac的修饰水平，增强SCNT胚胎的核重编程能力 Increase the levels of H3K9ac and H4K12ac in pseudoprokaryotic stage and enhance nuclear reprogramming in SCNT embryos	25.5 vs 10.7
Oxamflatin^［59］	1 µmol·L^-1	15 h	提高多能基因POU5F在囊胚阶段的表达和2-细胞和4-细胞期的组蛋白H3K9和H4K5乙酰化修饰水平，降低重构胚胎原核期去乙酰化酶活性和2-细胞期的DNA甲基化水平Increase expression of the pluripotency gene POU5F at the blastocyst stage and histone H3K9 and H4K5 acetylation modification levels at the 2-cell and 4-cell stages， and decrease deacetylase activity at the prokaryotic stage of remodeled embryos and DNA methylation levels at the 2-cell stage	23.6 vs 11.7

化合物 Compound	用量 Dosage	时间 Time	作用机制 Mechanism	囊胚率（处理vs对照） Blastocyst （treated vs control）/%
MC1568^［60］	10 µmol·L^-1	12 h	提高 SCNT 1-细胞和2-细胞 H4K12的乙酰化修饰水平；促进SCNT 囊胚OCT4、CDX2、SOX2和NANOG基因的表达 Increase the level of acetylase modification of H4K12 in SCNT 1-cell and 2-cell； promotes the expression of OCT4， CDX2， SOX2 and NANOG in SCNT blastocyst	（32.1±3.1） vs （21.1±1.4）
BIX-01294^［61］	50 nmol·L^-1	14~ 16 h	降低2-细胞和4-细胞阶段H3K9me2和H3K9me 的修饰水平，增加多能性基因SOX2，NANOG和OCT4的表达 Decrease the levels of H3K9me2 and H3K9me at 2-cell and 4-cell and increase the expression of SOX2， NANOG and OCT4	23.2 vs 16.4
毛壳素^［62］ Chaetocin	0.5 nmol·L^-1	24 h	纠正克隆胚胎异常的甲基化水平，调节表观遗传重编程和自噬活性 Correction of aberrant methylation levels and regulation of epigenetic reprogramming and autophagic activity in cloned embryos	（35.1±1.5） vs （24.3±1.1）
辛二酰苯胺异羟肟酸^［63］ SAHA	7. 5 µmol·L^-1	12 h	纠正克隆胚胎组蛋白乙酰化和相关基因表达的异常，使克隆胚胎H4K8ac水平、OCT4和HDAC1基因相对表达量趋近于IVF胚胎 Correction of abnormal histone acetylation and gene expression， improve the levels of H4K8ac， the expression of OCT4 and HDAC1 close to IVF embryos	33.9 vs 27.0
Sonic Hedgehog^［64］	1 µg·mL^-1	7 d	增加胚胎发育过程中的细胞增殖并减少凋亡 Increase cell proliferation and decreases apoptosis during embryonic development	50.3 vs 26.8
丙戊酸^［65］ Valproic acid	2 mmol·L^-1	24 h		（35.8±7.7） vs （14.7±6.6）
氨基酸^{［66‑67］} Amino acid	1%非必需氨基酸 1% non-essential amino acids	160~168 h		（32.2±1.0） vs （28.1±1.4）

化合物 Compound	用量 Dosage	时间 Time	作用机制 Mechanism	囊胚率（处理vs对照） Blastocyst （treated vs control）/%
MC1568^［60］	10 µmol·L^-1	12 h	提高 SCNT 1-细胞和2-细胞 H4K12的乙酰化修饰水平；促进SCNT 囊胚OCT4、CDX2、SOX2和NANOG基因的表达 Increase the level of acetylase modification of H4K12 in SCNT 1-cell and 2-cell； promotes the expression of OCT4， CDX2， SOX2 and NANOG in SCNT blastocyst	（32.1±3.1） vs （21.1±1.4）
BIX-01294^［61］	50 nmol·L^-1	14~ 16 h	降低2-细胞和4-细胞阶段H3K9me2和H3K9me 的修饰水平，增加多能性基因SOX2，NANOG和OCT4的表达 Decrease the levels of H3K9me2 and H3K9me at 2-cell and 4-cell and increase the expression of SOX2， NANOG and OCT4	23.2 vs 16.4
毛壳素^［62］ Chaetocin	0.5 nmol·L^-1	24 h	纠正克隆胚胎异常的甲基化水平，调节表观遗传重编程和自噬活性 Correction of aberrant methylation levels and regulation of epigenetic reprogramming and autophagic activity in cloned embryos	（35.1±1.5） vs （24.3±1.1）
辛二酰苯胺异羟肟酸^［63］ SAHA	7. 5 µmol·L^-1	12 h	纠正克隆胚胎组蛋白乙酰化和相关基因表达的异常，使克隆胚胎H4K8ac水平、OCT4和HDAC1基因相对表达量趋近于IVF胚胎 Correction of abnormal histone acetylation and gene expression， improve the levels of H4K8ac， the expression of OCT4 and HDAC1 close to IVF embryos	33.9 vs 27.0
Sonic Hedgehog^［64］	1 µg·mL^-1	7 d	增加胚胎发育过程中的细胞增殖并减少凋亡 Increase cell proliferation and decreases apoptosis during embryonic development	50.3 vs 26.8
丙戊酸^［65］ Valproic acid	2 mmol·L^-1	24 h		（35.8±7.7） vs （14.7±6.6）
氨基酸^{［66‑67］} Amino acid	1%非必需氨基酸 1% non-essential amino acids	160~168 h		（32.2±1.0） vs （28.1±1.4）

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提高猪体细胞克隆胚胎发育率的研究进展

Progress of the Improvement of Development of Porcine Somatic Cell Nuclear Transfer Embryos

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