3D培养在家畜卵母细胞及胚胎培养的研究现状与应用前景

doi:10.11843/j.issn.0366-6964.2025.04.003

摘要/Abstract

摘要：

三维(3D)细胞培养技术能够准确地模拟细胞的实际微环境，并再现细胞间的相互作用，使细胞行为特性更接近于生物体内的生存状态。因此，3D细胞培养技术广泛应用于家畜卵母细胞和胚胎发育等方面的研究，在很大程度上提高了体外卵母细胞和胚胎发育的质量，促进了研究结果的一致性和可靠性。本文通过材料、基质和方法3个方面论述了3D体外培养的发展，并基于细胞的生物学特性、培养条件、所使用的支架材料，细胞间的相互作用以及培养系统的设计总结了新的分类方法，即：细胞聚集系统、构建结构支架、构建仿生支架、与多学科交叉法(延伸)。在未来，通过对3D模型细胞培养技术的改进及优化，将为不同家畜卵母细胞体外培养和胚胎发育等方面的提供重要技术支持和新的可能性。

关键词: 三维培养系统, 卵母细胞, 体外胚胎

Abstract:

Three-dimensional (3D) cell culture technology can accurately simulate the actual microenvironment of cells and reproduce the interactions between cells, so that the behavior characteristics of cells are more similar to the living state in vivo. Therefore, 3D cell culture technology has been widely used in the research of oocyte and embryo development in domestic animals, which has greatly improved the quality of oocyte and embryo development in vitro, and thus promoted the consistency and reliability of research results. This paper discusses the development of 3D in vitro culture from three aspects: materials, substrates and methods, and summarizes new classification methods based on the biological characteristics of cells, culture conditions, scaffold materials used, interactions between cells and the design of culture systems, namely: cell aggregation system, construction of structural scaffolds, construction of bionic scaffolds, and multidisciplinary intersections (extension). In the future, through the improvement and optimization of 3D model cell culture technology, it will provide important technical support and new possibilities for the research of in vitro culture of different domestic oocytes and embryonic development.

Key words: three-dimensional culture system, oocytes, in vitro embryos

中图分类号:

S814.1

闫瑞, 贾朝阳, 马静, 杨娟, 刘新峰, 陈强. 3D培养在家畜卵母细胞及胚胎培养的研究现状与应用前景[J]. 畜牧兽医学报, 2025, 56(4): 1494-1507.

YAN Rui, JIA Chaoyang, MA Jing, YANG Juan, LIU Xinfeng, CHEN Qiang. The Research Status and Application Prospect of 3D Culture in Livestock Oocytes and Embryos Culture[J]. Acta Veterinaria et Zootechnica Sinica, 2025, 56(4): 1494-1507.

图/表 2

图 1

表 1

3D培养方法分类表"

方法Method	代表Represention	介绍Introduction	特点Characteristic	适用细胞Applicable cell
细胞聚集系统Cell aggregation system	悬滴培养板	将细胞包裹在悬滴中，在无接触的环境下细胞自聚集形成球状体。这种方法通常使用特殊设计的悬滴板架来创建包含单细胞悬液的小液滴，从而促进细胞的自组装和球形化	可以模拟细胞在体内的生长环境，增加细胞外基质的分泌，增强细胞之间的接触，更有利于激发细胞的生物活性可以实现多种细胞的共混培养，这对于研究细胞间的相互作用和信号传递具有重要意义	肿瘤细胞，干细胞，成纤维细胞等
构建结构支架Construct support structure	微载体	这种支架通常由生物兼容性良好的材料制成，如胶原蛋白、纤维素、黏多糖等天然聚合物，或者聚苯乙烯、二乙基氨基乙基葡聚糖等合成材料	能够提供一个更为接近细胞在体内生长状态的环境，有助于细胞之间的相互作用和信号传递这种支架的孔隙度、纤维性、渗透性和力学稳定性都可以被调节，以适应不同类型细胞的生长需求。可以促进细胞的生长和分化，提高细胞的活性和增殖能力	肿瘤细胞，干细胞，成纤维细胞，上皮细胞等
构建仿生支架Construction of biomimetic scaffolds	matrigel	使用matrigel这一基底膜基质材料。matrigel是从富含胞外基质蛋白的恩格尔布雷特-霍尔姆-斯沃肉瘤(Engelbreth-Holm-Swarm, EHS)小鼠肿瘤中提取的基底膜基质，其主要成分包括层黏连蛋白、Ⅳ型胶原、硫酸肝素糖蛋白、巢蛋白，还包含多种生长因子和基质金属蛋白酶等	能够较好地模拟体内细胞外基质的环境，有助于维持细胞的三维立体结构和自然的增殖分化活性。由于matrigel是动物来源的产品，可能存在批间差异和潜在的免疫反应问题	上皮细胞，内皮细胞，神经细胞，肌肉细胞，肿瘤细胞，干细胞等
	VitroGel	VitroGel是一种即用型、无外源性(Xeno-Free)的多糖水凝胶体系，它可以模拟自然的细胞外基质环境；VitroGel的使用非常方便，只需与细胞培养基混合即可转变为可调的水凝胶基质	即用型：单瓶体系，只需与细胞混匀即可；无异源蛋白成分：为无动物源性多糖水凝胶体系；室温稳定：该水凝胶体系室温稳定，pH中性；透明：该水凝胶体系颜色透明，兼容不同的成像系统，便于细胞观察；细胞容易收获：3D细胞培养后，可通过离心水凝胶中轻松收获细胞；可注射：使用恰当的混合比例，该水凝胶可用于注射，适合体内研究	诱导性多能干细胞或原代细胞，悬浮细胞，贴壁细胞，癌细胞，干细胞等
与多学科交叉法Disciplinary crossing and integration	微流控	利用微流控芯片来控制细胞培养基的流动，形成适合的剪切力、压力以及张力等，从而构建适合细胞生长的三维环境	可以模拟细胞在体内的微环境，包括气体、营养物质等物质的梯度浓度变化，以及细胞间的相互作用和细胞的生化和生理反应。微流控芯片的设计和制造需要精密的技术，成本相对较高。此外，微流控法的操作复杂，需要专业的设备和技术人员，这也限制其在一些研究领域中的应用	肿瘤细胞，干细胞，肝细胞，肾细胞等

表 1

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