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

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

• Technique and Method • Previous Articles Next Articles

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 Online:2025-11-24 Published:2025-06-26
Contact: Zicong Li E-mail:2393662931@qq.com;lizicong@scau.edu.cn
Supported by:
Seed Industry Revitalisation Project of the Guangdong Provincial Department of Agriculture and Rural Affairs(2024-XPY-00-004)

Abstract

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

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.

Figures/Tables 13

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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

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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