Xu Xiaolin, Guan Wencai, Yu Xiaolei, Xu Guoxiong, Wang Chenglong
Research Center for Clinical Medicine, Jinshan Hospital, Fudan University, Shanghai, 201508, P.R. China.
The State Key Laboratory of Metal Matrix Composites, School of Materials Science and Engineering, Shanghai Jiao Tong University, Shanghai, 200240, P. R. China.
Biomater Res. 2023 Jul 3;27(1):64. doi: 10.1186/s40824-023-00402-w.
Protocell refers to the basic unit of life and synthetic molecular assembly with cell structure and function. The protocells have great applications in the field of biomedical technology. Simulating the morphology and function of cells is the key to the preparation of protocells. However, some organic solvents used in the preparation process of protocells would damage the function of the bioactive substance. Perfluorocarbon, which has no toxic effect on bioactive substances, is an ideal solvent for protocell preparation. However, perfluorocarbon cannot be emulsified with water because of its inertia.
Spheroids can be formed in nature even without emulsification, since liquid can reshape the morphology of the solid phase through the scouring action, even if there is no stable interface between the two phases. Inspired by the formation of natural spheroids such as pebbles, we developed non-interfacial self-assembly (NISA) of microdroplets as a step toward synthetic protocells, in which the inert perfluorocarbon was utilized to reshape the hydrogel through the scouring action.
The synthetic protocells were successfully obtained by using NISA-based protocell techniques, with the morphology very similar to native cells. Then we simulated the cell transcription process in the synthetic protocell and used the protocell as an mRNA carrier to transfect 293T cells. The results showed that protocells delivered mRNAs, and successfully expressed proteins in 293T cells. Further, we used the NISA method to fabricate an artificial cell by extracting and reassembling the membrane, proteins, and genomes of ovarian cancer cells. The results showed that the recombination of tumor cells was successfully achieved with similar morphology as tumor cells. In addition, the synthetic protocell prepared by the NISA method was used to reverse cancer chemoresistance by restoring cellular calcium homeostasis, which verified the application value of the synthetic protocell as a drug carrier.
This synthetic protocell fabricated by the NISA method simulates the occurrence and development process of primitive life, which has great potential application value in mRNA vaccine, cancer immunotherapy, and drug delivery.
原细胞是指具有细胞结构和功能的生命基本单位及合成分子聚集体。原细胞在生物医学技术领域有巨大应用。模拟细胞的形态和功能是制备原细胞的关键。然而，原细胞制备过程中使用的一些有机溶剂会损害生物活性物质的功能。对生物活性物质无毒性作用的全氟碳是制备原细胞的理想溶剂。然而，全氟碳因其惰性无法与水乳化。
即使不进行乳化，自然界中也能形成球体，因为液体可通过冲刷作用重塑固相的形态，即便两相之间不存在稳定界面。受卵石等天然球体形成的启发，我们开发了微滴的非界面自组装（NISA）方法，作为迈向合成原细胞的一步，其中利用惰性全氟碳通过冲刷作用重塑水凝胶。
通过基于NISA的原细胞技术成功获得了合成原细胞，其形态与天然细胞非常相似。然后我们在合成原细胞中模拟细胞转录过程，并将原细胞用作mRNA载体转染293T细胞。结果表明原细胞递送了mRNA，并在293T细胞中成功表达了蛋白质。此外，我们使用NISA方法通过提取和重新组装卵巢癌细胞的膜、蛋白质和基因组来制造人工细胞。结果表明成功实现了肿瘤细胞的重组，其形态与肿瘤细胞相似。此外，通过NISA方法制备的合成原细胞用于通过恢复细胞钙稳态来逆转癌症化疗耐药性，这验证了合成原细胞作为药物载体的应用价值。
通过NISA方法制备的这种合成原细胞模拟了原始生命的发生和发展过程，在mRNA疫苗、癌症免疫治疗和药物递送方面具有巨大的潜在应用价值。
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