牛体外胚胎生产过程中培养液添加物研究进展

doi:10.11843/j.issn.0366-6964.2024.08.005

摘要/Abstract

摘要：

牛体外胚胎生产是一种广泛应用于畜牧生产和生物研究的现代繁殖技术，在促进肉牛和奶牛品种改良以及高效繁育中发挥着关键作用。随着分子生物学、生物化学和细胞生物学技术的不断发展，牛体外胚胎生产技术已取得了长足进步。然而，当前牛体外胚胎生产技术仍存在胚胎生产效率低和胚胎发育潜能差等瓶颈问题，在很大程度上限制了该技术的规模化推广应用。本文系统综述了牛体外胚胎生产中经典添加物和最新添加物的研究进展，旨在为优化构建不同添加物组合的技术体系提供参考，以期更加高效地获得牛体外优质胚胎。

关键词: 牛, 胚胎, 培养液, 受精, 添加物

Abstract:

Bovine embryo in vitro production (IVP) is a modern reproductive biotechnology, which is widely used in livestock production and biological research and plays an essential role in promoting breed improvement and reproductive efficiency for both beef and dairy cattle. As development of molecular biology, biochemistry and cell biology, a great progress has been made in bovine IVP. However, the current bovine IVP is still facing bottleneck problems, such as low efficiency of in intro embryo production and poor embryonic development potential, seriously hindering the further large-scale promotion and application of this technology. In this paper, the research progress of classical and latest additives in bovine IVP was systematically reviewed, intending to provide references for optimizing and constructing the technical system of different additive combinations, and aiming to more efficiently obtain high-quality bovine in vitro embryos.

Key words: bovine, embryo, culture solution, fertilization, additive

中图分类号:

S823.3

戴舒颖, 刘青, 李爱国, 余博, 陈洪波. 牛体外胚胎生产过程中培养液添加物研究进展[J]. 畜牧兽医学报, 2024, 55(8): 3309-3320.

Shuying DAI, Qing LIU, Aiguo LI, Bo YU, Hongbo CHEN. Research Progress on Culture Medium Additives in Bovine In Vitro Embryo Production[J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(8): 3309-3320.

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作者、年份 Author, Year	培养液 Culture solution	基础成分 Basis ingredient	添加物 Additive	卵母细胞成熟率/% Oocyte maturation rate
Ríos等^[32]，2014	M199	0.1 mg·mL^-1 L-glutamine、 2.2 mg·mL^-1 NaHCO3	10 ng·mL^-1L EGF、 15 mg·mL^-1 hyaluronic acid(HA)	75.80
Anchordoquy等^[33]，2015	TCM-199	10% FCS	300 ng·mL^-1 vascular endothelial growth factor (VEGF)	82.10±2.60
Canel等^[34]，2018	TCM-199	10% FBS、0.3 mmol pyruvate、2 μg·mL^-1 FSH	0.1 mmol cysteamine	82.73
Cajas等^[31]，2020	TCM-199	0% FCS、10 ng·mL^-1 EGF	25 μmol nobiletin	85.70±0.30
吉文汇等^[35]，2022	M199	10% FBS、1% FSH、 0.05% LH、0.2% E2	2 μmol维生素A	85.77±0.37
孙敬宇^[72]，2023	TCM-199	10 ng·mL^-1 EGF、0.04 mg·mL^-1 FSH、10% FBS、1% P-S、 0.022 g·mL^-1 pyruvate、 0.9 mg·mL^-1 cysteamine	20 μmol·L^-1柠檬苦素	80.13±2.01
Yang等^[36]，2022	M199	10 μg·mL^-1 E2	25 μg·mL^-1 connexin 37(Cx37)、50 ng·mL^-1 EGF、100 ng·mL^-1 IGF-1	87.43±6.86
El-Sheikh等^[29]，2023	M199	10 μg·mL^-1 FSH、1 μg·mL^-1 estradiol-17β、10 ng·mL^-1 EGF、 0.2 mmol sodium pyruvate、 0.6 mmol cysteine、0.1 mg·mL^-1 streptomycin	2 μmol vanillic acid (VA)	84.00
Wang等^[37]，2023	TCM-199	10% FBS、2 μg·mL^-1 estradiol、 10 ng·mL^-1 EGF、10 IU·mL^-1 FSH、10 IU·mL^-1 LH、 100 mmol pyruvate	5 μmol ferulic acid	71.20±3.838
Bicici等^[38]，2023	BO-IVM	/	0.5 mmol putrescine	74.21
Davoodian等^[39]，2023	M199	10% FBS、0.33 mmol sodium pyruvate、0.05 IU·mL^-1 FSH	5 μmol γ-oryzanol(ORY)	88.00±2.00

作者、年份 Author, Year	培养液 Culture solution	基础成分 Basis ingredient	添加物 Additive	卵裂率/% Cleavage rate
Gonçalves等^[57]，2013	TALP	0.2 mmol Na-pyruvate、6 mg·mL^-1 FAF-BSA、25 mmol sodium bicarbonate、 13 mmol Na-lactate、10 μg·mL^-1 heparin	10 μmol hypotaurine、 20 μmol penicillamine、 2 μmol epinephrine	70.70
Santana等^[58]， 2016	IVF	1 mmol sodium pyruvate、21.6 mmol sodium lactate、6 mg·mL^-1 BSA、 50 mg·mL^-1 gentamicin、2 mmol penicillamine、1 mmol hypotaurine、 250 mmol epinephrine	10 mmol L-arginine (ARG)	83.90±0.90
Kang等^[59]， 2015	mBO	/	2.5 mmol theophylline、 20 μmol d-penicillamine、 10 μmol hypotaurine、 1 μmol epinephrine	86.20±8.80
An等^[60]，2017	BO	/	40 μg·mL^-1 heparin	66.70±1.50
Wrenzycki等^[45]，2019	TALP	2 mmol penicillamine、1 mmol hypotaurine、250 mmol epinephrine	10 μg·mL^-1 heparin	82.40
Yuan等^[61]， 2020	IVF	6 mg·mL^-1 BSA、22 mg·mL^-1 sodium pyruvate、100 IU·mL^-1 penicillin、0.1 mg·mL^-1 streptomycin	10 mmol nicotinamide	80.00
Rodriguez- villarnil等^[62]， 2020	Fert-TALP	/	5 μg·mL^-1 heparin、 5 μg·mL^-1 binder of sperm protein 1 (BSP1)	76.40±2.80
Zhang等^[63]， 2023	/	/	100 nmol c-type natriuretic peptide (CNP)、10~9mol melatonin (MT)	91.61±5.17

作者、年份 Author, Year	培养液 Culture solution	基础成分 Basis ingredient	添加物 Additive	囊胚率/% Blastocyst rate
Mesalam等^[67]，2019	SOF-BE1	4 mg·mL^-1 BSA	100 ng·mL^-1 EGF、5mg·mL^-1 insulin、5 mg·mL^-1 transferrin、 5 ng·mL^-1 sodium selenite	41.42±0.70
Zolini等^[76]，2019	SOF-BE2	3.7 mg·mL^-1 EFAF-BSA	1.5 mmol L-carnitine	23.60±1.70
El-Maarri等^[77], 2019	SOF	EFAF-BSA	10 ng·mL^-1 EGF、1 mg·mL^-1 hyaluronic acid(HA)	39.70±7.40
Anchordoquy等^[28]，2022	SOFm	/	6 ng·mL^-1 Mn、100 ng·mL^-1Se、 200 ng·mL^-1 Cu、400 ng·mL^-1L Zn	46.06
Fabra等^[78]，2022	SOF	1 mmol glutamine、2% MEM-EAA、1% MEM-NEAA、 8 mg·mL^-1BSA-FAF	2.5 μmol alpha-lipoic acid	29.79
Miglio等^[79]，2023	CR2	50 mg·mL^-1amikacin、 0.1 mmol amino acids、 2.5% FBS、 6 mg·mL^-1BSA	3 μmol ZnO(np)+CUR(姜黄素功能化氧化锌纳米颗粒)	22.1±6.61
El-Sheikh等^[29]，2023	IVC medium	/	2 μmol vanillic acid (VA)	38.75±2.87

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