Huang Aria Zheyuan, Prahl Louis S, Xu Karen, Mauck Robert L, Burdick Jason A, Hughes Alex J
Department of Bioengineering, University of Pennsylvania, Philadelphia, PA, USA.
Bioengineering Graduate Group, University of Pennsylvania, Philadelphia, PA, USA.
Nat Commun. 2025 Aug 22;16(1):7829. doi: 10.1038/s41467-025-63197-5.
Kidney explants are traditionally cultured at air-liquid interfaces, which disrupts 3D tissue structure and limits interpretation of developmental data. Here we develop a 3D culture technique using hydrogel embedding to capture kidney morphogenesis in real time. 3D culture better approximates in vivo-like niche spacing and tubule dynamics, as well as branching defects under control conditions and GDNF-RET signaling perturbations. To isolate the effect of material properties on explant development, we apply acrylated hyaluronic acid hydrogels that allow independent tuning of stiffness and adhesion. We find that sufficient stiffness and adhesive ligands are both required to maintain kidney shape. More adhesive hydrogels increase nephrons per ureteric bud (UB) tip while matrix stiffness has a "Goldilocks effect" centered at ~2 kPa. Our technique captures large-scale, in vivo-like tissue morphogenesis in 3D, improving insight into congenital disease phenotypes. Moreover, understanding the impact of boundary condition mechanics on kidney development benefits fundamental research and renal engineering.
传统上，肾外植体是在气液界面进行培养的，这会破坏三维组织结构并限制对发育数据的解读。在此，我们开发了一种使用水凝胶包埋的三维培养技术，以实时捕捉肾脏形态发生过程。三维培养能更好地模拟体内样的微环境间距和肾小管动态，以及在对照条件和GDNF-RET信号扰动下的分支缺陷。为了分离材料特性对外植体发育的影响，我们应用了丙烯酸化透明质酸水凝胶，其允许独立调节硬度和黏附性。我们发现，维持肾脏形状既需要足够的硬度，也需要黏附配体。黏附性更强的水凝胶会增加每个输尿管芽（UB）尖端的肾单位数量，而基质硬度具有以约2kPa为中心的“金发姑娘效应”。我们的技术在三维空间中捕捉大规模的、类似体内的组织形态发生，有助于深入了解先天性疾病表型。此外，了解边界条件力学对肾脏发育的影响有利于基础研究和肾脏工程。
