改变表观遗传修饰和供体细胞代谢状态可提高犬-猪异种克隆胚胎的发育效率

doi:10.16288/j.yczz.25-109

遗传 ›› 2025, Vol. 47 ›› Issue (12): 1365-1376.doi: 10.16288/j.yczz.25-109

改变表观遗传修饰和供体细胞代谢状态可提高犬-猪异种克隆胚胎的发育效率

闫艳霞¹^,²(), 张雨涵¹^,², 吴珍芳¹^,², 李紫聪¹^,²()

1.华南农业大学动物科学学院，广州 510642
2.国家生猪种业工程技术研究中心，广州 510642

收稿日期:2025-04-17 修回日期:2025-05-27 出版日期:2025-11-24 发布日期:2025-06-26
通讯作者: 李紫聪，博士，教授，研究方向：动物体细胞克隆技术和克隆胚胎发育调控机制。E-mail: lizicong@scau.edu.cn
作者简介:闫艳霞，硕士研究生，专业方向：畜牧学。E-mail：2393662931@qq.com
基金资助:
广东省农业农村厅种业振兴项目(2024-XPY-00-004)

Changes of epigenetic modification and donor cell metabolic status can improve the developmental efficiency of canine-porcine interspecies somatic cell nuclear transfer embryos

Yanxia Yan¹^,²(), Yuhan Zhang¹^,², Zhenfang Wu¹^,², Zicong Li¹^,²()

1. College of Animal Science, South China Agricultural University, Guangzhou 510642, China
2. National Engineering Research Center for Breeding Swine Industry, Guangzhou 510642, China

Received:2025-04-17 Revised:2025-05-27 Published:2025-11-24 Online:2025-06-26
Supported by:
Seed Industry Revitalisation Project of the Guangdong Provincial Department of Agriculture and Rural Affairs(2024-XPY-00-004)

摘要/Abstract

摘要：

为了探索能提高犬体细胞克隆效率的新方法，本研究比较了糖酵解促进剂PS48和表观遗传修饰剂(DNA甲基化酶抑制剂RG108和组蛋白去乙酰化酶抑制剂Scriptaid)的不同处理浓度和时间对犬-猪异种体细胞核移植(interspecies somatic cell nuclear transfer，iSCNT)胚胎发育效率的影响。结果显示：(1) 5 μmol/L PS48处理犬耳成纤维细胞(canine ear fibroblasts，cEFs)和犬脂肪间充质干细胞(canine adipose tissue-derived mesenchymal stem cells，cAd-MSCs) 24 h显著增强了其后续iSCNT胚胎的发育能力，PS48处理cEFs组的iSCNT胚胎的卵裂率、4-细胞期率和8-细胞期率均显著高于对照组(46.90±1.64% vs 13.30±1.61%，32.30±1.55% vs 8.26±0.88%，10.62±1.68% vs 5.50±0.84%；P<0.05)，PS48处理cAd-MSCs组的iSCNT胚胎的卵裂率和4-细胞期率显著高于对照组(49.51±3.00% vs 31.25±2.73%，26.21±2.08% vs 15.18±1.58%；P<0.05)；(2) 20 μmol/L RG108处理cEFs和cAd-MSCs 48 h对iSCNT胚胎发育效率无显著影响，0 nmol/L、400 nmol/L、500 nmol/L和600 nmol/L Scriptaid分别处理cEFs和cAd-MSCs 24 h对iSCNT胚胎发育效率无显著影响；(3) 20 μmol/L RG108处理两种供体细胞来源iSCNT胚胎对其发育有显著促进作用(P<0.05)，500 nmol/L Scriptaid处理cEFs来源的iSCNT胚胎16 h可显著提高其卵裂率和4-细胞期率(23.08±2.94% vs 9.47±1.70%，18.68±3.25% vs 6.32±1.07%；P<0.05)。本研究建立了能显著提高犬-猪iSCNT胚胎发育效率的新方法，有助于促进犬体细胞克隆技术的应用和发展。

关键词: 供体细胞代谢, 表观遗传修饰, 犬克隆, 体细胞核移植, PS48, RG108, Scriptaid

Abstract:

To explore new ways to improve canine cloning efficiency, this study compared the effects of different concentrations and treatment times of glycolysis promoter PS48 and epigenetic modifiers (DNA methylase inhibitor RG108 and histone deacetylase inhibitor Scriptaid) on the developmental ability of canine-porcine interspecies somatic cell nuclear transfer (iSCNT) embryos. The results showed that (1) 5 μmol/L PS48 treatment on canine ear fibroblasts (cEFs) and canine adipose tissue-derived mesenchymal stem cells (cAd-MSCs) for 24 h significantly enhanced subsequent iSCNT embryo development. The cleavage rate, 4-cell stage rate and 8-cell stage rate of iSCNT embryos produced from PS48-treated cEFs were significantly higher than those of control iSCNT embryos (46.90±1.64% vs 13.30±1.61%, 32.30±1.55% vs 8.26±0.88%, and 10.62±1.68% vs 5.50±0.84%; P<0.05). The cleavage and 4-cell stage rates of iSCNT embryos generated from PS48-treated cAd-MSCs were significantly higher than those of control iSCNT embryos (49.51±3.00% vs 31.25±2.73%, 26.21±2.08% vs 15.18±1.58%; P<0.05). (2) Treatment of cEFs and cAd-MSCs with 20 μmol/L RG108 for 48 h had no significant effect on the developmental efficiency of iSCNT embryos. Treatment of cEFs and cAd-MSCs with 0 nmol/L, 400 nmol/L, 500 nmol/L and 600 nmol/L Scriptaid for 24 h had no significant effect on the developmental efficiency of iSCNT embryos. (3) Treatment of iSCNT embryos derived from two types of donor cells with 20 μmol/L RG108 significantly promoted their developmental competence (P<0.05). Treatment of iSCNT embryos derived from cEFs with 500 nmol/L Scriptaid for 16 h significantly increased their cleavage and 4-cell stage rates (23.08±2.94% vs 9.47±1.70%, 18.68±3.25% vs 6.32±1.07%; P<0.05). This study established some new methods that can significantly improve the developmental efficiency of canine-porcine iSCNT embryos, thereby contributing to the development and application of canine somatic cell cloning technology.

Key words: donor cell metabolism, epigenetic modifications, canine cloning, somatic cell nuclear transfer (SCNT), PS48, RG108, Scriptaid

闫艳霞, 张雨涵, 吴珍芳, 李紫聪. 改变表观遗传修饰和供体细胞代谢状态可提高犬-猪异种克隆胚胎的发育效率[J]. 遗传, 2025, 47(12): 1365-1376.

Yanxia Yan, Yuhan Zhang, Zhenfang Wu, Zicong Li. Changes of epigenetic modification and donor cell metabolic status can improve the developmental efficiency of canine-porcine interspecies somatic cell nuclear transfer embryos[J]. Hereditas(Beijing), 2025, 47(12): 1365-1376.

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组别	克隆胚胎数(n)	卵裂率(%)	4-细胞期率(%)	8-细胞期率(%)	16-细胞期率(%)
对照组	218	13.30^a±1.61	8.26^a±0.88	5.50^a±0.84	1.38^a±0.93
PS48处理组	226	46.90^b±1.64	32.30^b±1.55	10.62^b±1.68	3.54^a±0.52

组别	克隆胚胎数(n)	卵裂率(%)	4-细胞期率(%)	8-细胞期率(%)	16-细胞期率(%)
对照组	112	31.25^a±2.73	15.18^a±1.58	4.46^a±0.52	2.68^a±0.04
PS48处理组	103	49.51^b±3.00	26.21^b±2.08	4.85^a±0.46	4.85^a±0.46

组别	克隆胚胎数(n)	卵裂率(%)	4-细胞期率(%)	8-细胞期率(%)	16-细胞期率(%)
对照组	81	17.28^a±1.70	11.11^a±0.99	6.17^a±2.03	2.47^a±2.38
RG108处理组	76	7.89^a±1.26	7.89^a±1.36	6.58^a±2.23	5.26^a±1.89

组别	克隆胚胎数(n)	卵裂率(%)	4-细胞期率(%)	8-细胞期率(%)	16-细胞期率(%)
对照组	150	12.00^a±1.33	8.00^a±1.25	3.33^a±0.42	2.67^a±0.47
RG108处理组	141	9.22^a±0.28	3.55^a±0.90	2.13^a±1.00	1.42^a±1.42

处理浓度(nmol/L)	克隆胚胎数(n)	卵裂率(%)	4-细胞期率(%)	8-细胞期率(%)	16-细胞期率(%)
0	95	23.2^a±1.95	12.6^a±0.81	4.2^a±1.64	0^a±0
400	114	25.4^a±4.22	12.3^a±1.79	3.5^a±0.52	1.8^a±0.72
500	100	25.0^a±2.82	12.0^a±2.71	6.0^a±2.09	3.0^a±1.93
600	122	17.2^a±1.03	8.2^a±1.45	4.1^a±2.84	2.5^a±4.08

改变表观遗传修饰和供体细胞代谢状态可提高犬-猪异种克隆胚胎的发育效率

Changes of epigenetic modification and donor cell metabolic status can improve the developmental efficiency of canine-porcine interspecies somatic cell nuclear transfer embryos

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处理浓度(nmol/L)	克隆胚胎数(n)	卵裂率(%)	4-细胞期率(%)	8-细胞期率(%)	16-细胞期率(%)
0	117	27.4^a±0.39	17.9^a±0.45	9.4^ab±1.79	6.0^a±1.37
400	82	26.8^a±4.80	15.9^a±3.77	2.4^b±4.99	1.2^a±1.51
500	119	31.9^a±1.96	24.4^a±1.58	8.4^ab±1.69	5.0^a±2.66
600	108	35.2^a±1.89	21.3^a±1.45	11.1^a±1.39	6.5^a±0.57

组别	克隆胚胎数(n)	卵裂率(%)	4-细胞期率(%)	8-细胞期率(%)	16-细胞期率(%)
对照组	76	14.47^a±1.49	6.58^a±1.70	1.32^a±1.89	0^a±0
RG108处理组	74	31.08^b±1.07	21.62^b±1.01	12.16^b±1.18	6.76^b±0.88

组别	克隆胚胎数(n)	卵裂率(%)	4-细胞期率(%)	8-细胞期率(%)	16-细胞期率(%)
对照组	121	10.74^a±1.22	6.61^a±1.52	3.31^a±1.07	0.83^a±1.18
RG108处理组	125	21.60^b±1.45	8.00^a±1.34	3.20^a±1.14	2.40^a±1.18

组别	克隆胚胎数(n)	卵裂率(%)	4-细胞期率(%)	8-细胞期率(%)	16-细胞期率(%)
对照组	95	9.47^a±1.70	6.32^a±1.07	5.26^a±0.85	1.05^a±1.41
Scriptaid处理组	91	23.08^b±2.94	18.68^b±3.25	9.89^a±1.95	3.29^a±3.59

组别	克隆胚胎数(n)	卵裂率(%)	4-细胞期率(%)	8-细胞期率(%)	16-细胞期率(%)
对照组	120	60.8^a±3.31	32.5^a±2.55	5.0^a±1.44	5.0^a±1.44
Scriptaid处理组	116	24.1^b±3.75	9.5^b±2.1	4.3^a±1.44	2.6^a±1.27

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