乳液与微流控工艺在构建仿生细胞中的应用

doi:10.11949/0438-1157.20251141

摘要/Abstract

摘要：

乳液法与微流控技术在仿生细胞构建中具有关键作用和广阔应用前景。乳液法（包括 Pickering 乳液）以操作简便、设备依赖低及易于宏量制备的优势，成为封装酶、药物等功能组分的高效平台，尤其适用于对尺寸均一性要求不高的大规模应用。微流控技术则利用微米级通道精确操控流体，能够生成尺寸高度均一、结构可控的液滴，实现仿生细胞组成、尺寸与功能的精确编程，是高通量筛选、基础研究及定制化人工细胞构建的理想工具。两者互为补充，共同推动了仿生细胞在限域催化、底物筛选、信号传递及药物递送等领域的创新应用。尽管在构建精度、安全性和功能复杂性方面仍存在挑战，但随着多学科交叉发展，基于乳液与微流控的仿生细胞已成为探索生命本质与开发创新生物技术的强大平台。

关键词: 微流控, 液滴, 合成, 微反应器, 膜, 酶, 细胞工程

Abstract:

Emulsion-based methods and microfluidic technology play critical roles and demonstrate broad application prospects in the construction of biomimetic cells. Emulsion techniques (including Pickering emulsions) serve as efficient platforms for encapsulating functional components such as enzymes and drugs, benefiting from their operational simplicity, low equipment dependency, and ease of large-scale preparation. They are particularly suitable for mass production where high size uniformity is not a strict requirement. In contrast, microfluidics utilizes micron-scale channels to precisely manipulate fluids, generating highly monodisperse droplets with controllable structures. This enables precise programming of biomimetic cell composition, size, and function, making it an ideal tool for high-throughput screening, fundamental research, and customized artificial cell construction. These two approaches complement each other and jointly drive innovative applications of biomimetic cells in confined catalysis, substrate screening, signal transduction, and drug delivery. Although challenges remain in construction precision, safety, and functional complexity, the ongoing interdisciplinary convergence has established emulsion- and microfluidics-based biomimetic cells as powerful platforms for exploring the essence of life and developing innovative biotechnologies.

Key words: microfluidics, droplet, synthesis, microreactor, membranes, enzyme, cell engineering

中图分类号:

TQ464.8

周骏, 金烁, 王思远, 高冲, 陈笑非, 贺志远. 乳液与微流控工艺在构建仿生细胞中的应用[J]. 化工学报, DOI: 10.11949/0438-1157.20251141.

Jun ZHOU, Shuo JIN, Siyuan WANG, Chong GAO, Xiaofei CHEN, Zhiyuan HE. Processes and applications of emulsions and microfluidics in biomimetic cell construction[J]. CIESC Journal, DOI: 10.11949/0438-1157.20251141.

图/表 11

图1 采用油包水乳液法制备封装核结构的仿生细胞巨型单层囊泡（GUVs）[44]

Fig.1 Biomimetic cells with nuclear structures encapsulated within giant unilamellar vesicles (GUVs) via water-in-oil (W/O) emulsion method[44]

图2 基于皮克林乳液的含交联外壳的COF基多孔微反应器（CALB@COF-MCs-SH）的制备[51]

Fig.2 Fabrication of COF-based porous microreactors with cross-linked shells (CALB@COF-MCs-SH) via Pickering emulsion[51]

图3 微流控芯片及液滴产生过程：（a）T字型微流控芯片液滴生成示意图；（b）流动聚焦型微流控芯片液滴生成示意图；（c）微流控产生的分散均匀的微液滴[57-60]

Fig.3 Microfluidic Chip and Droplet Generation Process: (a) Schematic diagram of droplet generation in a T-junction microfluidic chip; (b) Schematic diagram of droplet generation in a flow-focusing microfluidic chip; (c) Uniformly dispersed microdroplets produced by microfluidics [57-60]

图4 微流控设备的尺寸和内外相流体流速对液滴尺寸的影响：(a) 平均直径为356.59μm(a1)、465.43μm(a2)、552.38μm(a3)、610.54μm(a4)、636.16μm(a5)和691.54μm(a6)的单分散液滴的光学显微照片；(b) 外相与内相的流速比(Uu0/ui)和微流控设备的尺寸对液滴尺寸的影响；(c) 不同平均尺寸液滴的变异系数(CV)值；(d) 液滴尺寸与微流控设备尺寸、连续相和分散相液体的粘度及流速之间的关系[61]

Fig.4 Effects of microfluidic device dimensions and fluid flow rates of both continuous and dispersed phases on droplet size:(a) Optical micrographs of monodisperse droplets with average diameters of 356.59 μm (a1), 465.43 μm (a2), 552.38 μm (a3), 610.54 μm (a4), 636.16 μm (a5), and 691.54 μm (a6).;(b) Influence of flow rate ratio (Uu0/ui) between continuous and dispersed phases and microfluidic device dimensions on droplet size;(c) Coefficient of variation (CV) values of droplets with different average sizes;(d) Relationship between droplet size and microfluidic device dimensions, viscosities and flow rates of continuous and dispersed phases[61]

图5 双乳液形成过程：(a) 示意图和 (b) 高速摄像显微镜图[62]

Fig.5 Formation Process of Double Emulsion: (a) Schematic diagram; (b) High-speed microscopic image [62]

图6 不同外部流量下乳液类型的相图:（a）S2G8（b）S4G6（c）S6G4以及（d）S8G2，其中×-不稳定乳液，■-多核乳液，●-单核乳液和⚪-非核乳液；（e）不同类型乳液的图像：I - 多核乳液，II - 单核乳液，III - 非核乳液，IV - 不稳定乳液[63]

Fig.6 Phase diagram of emulsion types under different external flow rates: (a) S2G8, (b) S4G6, (c) S6G4, and (d) S8G2, where ×- unstable emulsion, ■- multi-core emulsion, ●- single-core emulsion, and ⚪- non-nucleated emulsion; (e) Images of different emulsion types: I - multi-core emulsion, II - single-core emulsion, III - non-nucleated emulsion, IV - unstable emulsion[63]

图7 含海藻糖的聚合物囊泡纳米反应器形成过程示意图[67]

Fig.7 Formation of polymer vesicle nanoreactors containing trehalose[67]

图8 酶-MOF复合材料的微流控层流合成和本体溶液合成示意图[75]

Fig.8 Schematic illustration of microfluidic laminar flow synthesis versus bulk solution synthesis for enzyme-MOF composites[75]

图9 膜选择性示意图[60]

Fig.9 Schematic diagram of membrane selectivity[60]

图10 囊泡制备及信号传递：（a）通过微流控方法制备水包油包水（W1/O/W2）液滴；（b）由Pluronic L121组成的产生的聚合物体的示意图。PEG：聚乙二醇，PPG：聚丙二醇；（c）通过引入丙酮酸激酶（PyK）和乳酸脱氢酶（LDH）在聚合物体中驱动的PEP级联酶促反应的示意图[60]

Fig.10 Capsule Preparation and Signal Transduction: (a) Preparation of water-in-oil-in-water (W1/O/W2) double emulsion droplets via microfluidics; (b) Schematic diagram of the generated polymersomes composed of Pluronic L121. PEG: Polyethylene glycol, PPG: Polypropylene glycol; (c) Schematic illustration of the PEP-driven cascade enzymatic reaction in polymersomes through the introduction of pyruvate kinase (PyK) and lactate dehydrogenase (LDH) [60]

图11 模拟植物体中的双触发释放机制及具体性能 (a) 载药尺寸响应型两步释放机制示意图；(b) 体外释放研究装置的示意图；(c) 装载有4-或2000-kDa FITC-右旋糖酐的植物体的顺序释放曲线；(d) FITC-右旋糖酐负载的植物体在释放研究前后的照片；(e) 和(f) 4-kDa与2000-kDa FITC-葡聚糖尺寸分布的动态光散射（DLS）表征结果[92]

Fig.11 Simulation of Dual-Triggered Release Mechanism and Specific Performance in Phytomimetic Systems：(a) Schematic diagram of the cargo size-dependent two-step release mechanism; (b) Illustration of the in vitro release study setup.;(c) Sequential release profiles of 4-kDa or 2000-kDa FITC-dextran loaded phytosomes.;(d) Photographs of FITC-dextran loaded phytosomes before and after the release study; (e, f) DLS measurements showing the size distribution of 4-kDa and 2000-kDa FITC-dextran[92]

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