Stem Cell Research Lab, Department of Chemistry, Faculty of Science, Ankara University, Ankara 06560, Turkey.
School of Life Sciences, Faculty of Medicine and Health Sciences, University of Nottingham, Nottingham NG7 2UH, UK.
Acta Biomater. 2023 Nov;171:223-238. doi: 10.1016/j.actbio.2023.09.040. Epub 2023 Oct 2.
Organoids are an emerging technology with great potential in human disease modelling, drug development, diagnosis, tissue engineering, and regenerative medicine. Organoids as 3D-tissue culture systems have gained special attention in the past decades due to their ability to faithfully recapitulate the complexity of organ-specific tissues. Despite considerable successes in culturing physiologically relevant organoids, their real-life applications are currently limited by challenges such as scarcity of an appropriate biomimetic matrix. Peptide amphiphiles (PAs) due to their well-defined chemistry, tunable bioactivity, and extracellular matrix (ECM)-like nanofibrous architecture represent an attractive material scaffold for organoids development. Using cerebral organoids (COs) as exemplar, we demonstrate the possibility to create bio-instructive hydrogels with tunable stiffness ranging from 0.69 kPa to 2.24 kPa to culture and induce COs growth. We used orthogonal chemistry involving oxidative coupling and supramolecular interactions to create two-component hydrogels integrating the bio-instructive activity and ECM-like nanofibrous architecture of a laminin-mimetic PAs (IKVAV-PA) and tunable crosslinking density of hyaluronic acid functionalized with tyramine (HA-Try). Multi-omics technology including transcriptomics, proteomics, and metabolomics reveals the induction and growth of COs in soft HA-Tyr hydrogels containing PA-IKVAV such that the COs display morphology and biomolecular signatures similar to those grown in Matrigel scaffolds. Our materials hold great promise as a safe synthetic ECM for COs induction and growth. Our approach represents a well-defined alternative to animal-derived matrices for the culture of COs and might expand the applicability of organoids in basic and clinical research. STATEMENT OF SIGNIFICANCE: Synthetic bio-instructive materials which display tissue-specific functionality and nanoscale architecture of the native extracellular matrix are attractive matrices for organoids development. These synthetic matrices are chemically defined and animal-free compared to current gold standard matrices such as Matrigel. Here, we developed hydrogel matrices with tunable stiffness, which incorporate laminin-mimetic peptide amphiphiles to grow and expand cerebral organoids. Using multi-omics tools, the present study provides exciting data on the effects of neuro-inductive cues on the biomolecular profiles of brain organoids.
类器官是一种具有巨大潜力的新兴技术，可用于人类疾病建模、药物开发、诊断、组织工程和再生医学。由于类器官 3D 组织培养系统能够忠实地再现特定组织的复杂性，因此在过去几十年中得到了特别关注。尽管在培养具有生理相关性的类器官方面取得了相当大的成功，但由于缺乏合适的仿生基质等挑战，其实际应用目前受到限制。肽两亲物（PAs）由于其明确的化学性质、可调节的生物活性和细胞外基质（ECM）样纳米纤维结构，代表了开发类器官的有吸引力的材料支架。本文以脑类器官（COs）为例，证明了使用具有 0.69 kPa 至 2.24 kPa 可调硬度的生物指令性水凝胶来培养和诱导 COs 生长的可能性。我们使用涉及氧化偶联和超分子相互作用的正交化学方法，创建了两种组分的水凝胶，将层粘连蛋白模拟肽（IKVAV-PA）的生物指令性活性和 ECM 样纳米纤维结构以及通过 tyramine 官能化的透明质酸（HA-Try）的可调交联密度集成在一起。包括转录组学、蛋白质组学和代谢组学在内的多组学技术揭示了软质 HA-Tyr 水凝胶中包含 PA-IKVAV 的 COs 的诱导和生长，使得 COs 表现出与在 Matrigel 支架中生长的相似的形态和生物分子特征。我们的材料作为 COs 诱导和生长的安全合成 ECM 具有很大的应用前景。我们的方法代表了一种用于 COs 培养的明确替代动物衍生基质的方法，可能会扩大类器官在基础和临床研究中的应用。意义声明：具有组织特异性功能和天然细胞外基质纳米结构的合成生物指令性材料是类器官开发的有吸引力的基质。与当前的金标准基质（如 Matrigel）相比，这些合成基质具有化学定义性且不含动物。在这里，我们开发了具有可调硬度的水凝胶基质，其中包含层粘连蛋白模拟肽两亲物以生长和扩展大脑类器官。使用多组学工具，本研究提供了关于神经诱导线索对脑类器官生物分子特征影响的令人兴奋的数据。
