Rovers Maritza M, Slootweg Erik J, Los Ferdinand C O, Dankers Patricia Y W
Institute for Complex Molecular Systems, Eindhoven University of Technology, P.O. Box 513, Eindhoven, 5600 MB, The Netherlands.
Department of Biomedical Engineering, Laboratory of Chemical Biology, Eindhoven University of Technology, P.O. Box 513, Eindhoven, 5600 MB, The Netherlands.
Adv Biol (Weinh). 2025 Jun 4:e00690. doi: 10.1002/adbi.202400690.
Protoplast regeneration into plant cells and further into plants is an ongoing challenge in agricultural biotechnology. Inspired by mammalian tissue engineering, a strategic shift is proposed in plant tissue engineering to steer protoplast culture using fully synthetic materials-based culture platforms. Here a supramolecular materials method to engineer modular culture methods for protoplasts is chosen to use. Supramolecular monomers as modular building blocks allow to make various hydrogel formulations and to study different protoplast cultures; including 2D cultures on top of supramolecular hydrogels, 2.5D cultures using supramolecular fibers in solution, and 3D cultures when encapsulated in bulk hydrogels or microgels. Importantly, the need is shown for bioactive functionalization of the supramolecular hydrogels with a peptide additive in 2D protoplast cultures. After 11 days, the bioactive hydrogel induced protoplast enlargement, which is absent on pristine hydrogels. The opposite effect is present for protoplasts cultured in 3D, showing plasmolysis as a result of the bioactive additive. Interestingly, in 2.5D lower bioactive additive concentrations in supramolecular fibers stimulated protoplast enlargement, demonstrated by similar morphological changes as in 2D. Finally, protoplast encapsulation in supramolecular microgels is showcased. This work demonstrates the potential to modularly engineer various synthetic platforms to facilitate cellular agriculture.
原生质体再生为植物细胞并进一步再生为植物，是农业生物技术中一项持续存在的挑战。受哺乳动物组织工程的启发，植物组织工程领域提出了一项战略转变，即利用基于全合成材料的培养平台来引导原生质体培养。本文选择了一种超分子材料方法来设计用于原生质体的模块化培养方法。超分子单体作为模块化构建块，可用于制备各种水凝胶配方并研究不同的原生质体培养方式；包括在超分子水凝胶顶部进行的二维培养、在溶液中使用超分子纤维进行的2.5维培养，以及封装在块状水凝胶或微凝胶中的三维培养。重要的是，研究表明在二维原生质体培养中，需要用肽添加剂对超分子水凝胶进行生物活性功能化。11天后，生物活性水凝胶诱导原生质体增大，而在原始水凝胶上则未出现这种现象。在三维培养的原生质体中则出现相反的效果，即生物活性添加剂导致质壁分离。有趣的是，在2.5维培养中，超分子纤维中较低浓度的生物活性添加剂刺激了原生质体增大，其形态变化与二维培养相似。最后，展示了原生质体封装在超分子微凝胶中的情况。这项工作证明了模块化设计各种合成平台以促进细胞农业发展的潜力。
