3B's Research Group, I3Bs-Research Institute on Biomaterials, Biodegradables and Biomimetics, University of Minho, Headquarters of the European Institute of Excellence on Tissue Engineering and Regenerative Medicine, AvePark, Parque de Ciência e Tecnologia, Zona Industrial da Gandra, 4805-017 Barco, Guimarães, Portugal.
ICVS/3B's-PT Government Associate Laboratory, Braga/Guimarães, Portugal.
Biofabrication. 2021 Jul 26;13(4). doi: 10.1088/1758-5090/ac0fca.
Decellularized extracellular matrices (ECMs) are able to provide the necessary and specific cues for remodeling and maturation of tissue-specific cells. Nevertheless, their use for typical biofabrication applications requires chemical modification or mixing with other polymers, mainly due to the limited viscoelastic properties. In this study, we hypothesize that a bioink exclusively based on decellularized kidney ECM (dKECM) could be used to bioprint renal progenitor cells. To address these aims, porcine kidneys were decellularized, lyophilized and digested to yield a viscous solution. Then, the bioprinting process was optimized using an agarose microparticle support bath containing transglutaminase for enzymatic crosslinking of the dKECM. This methodology was highly effective to obtain constructs with good printing resolution and high structural integrity. Moreover, the encapsulation of primary renal progenitor cells resulted in high cell viability, with creation of 3D complex structures over time. More importantly, this tissue-specific matrix was also able to influence cellular growth and differentiation over time. Taken together, these results demonstrate that unmodified dKECM bioinks have great potential for bioengineering renal tissue analogs with promising translational applications and/or formodel systems. Ultimately, this strategy may have greater implications on the biomedical field for the development of bioengineered substitutes using decellularized matrices from other tissues.
去细胞细胞外基质（ECM）能够为组织特异性细胞的重塑和成熟提供必要的和特定的线索。然而，它们在典型的生物制造应用中的使用需要化学修饰或与其他聚合物混合，主要是由于其有限的粘弹性。在本研究中，我们假设仅基于去细胞肾脏 ECM（dKECM）的生物墨水可用于生物打印肾脏祖细胞。为了达到这些目的，对猪肾进行去细胞化、冻干和消化，以得到粘性溶液。然后，使用含有转谷氨酰胺酶的琼脂糖微球支持浴来优化生物打印过程，以实现 dKECM 的酶交联。这种方法非常有效，可以获得具有良好打印分辨率和高结构完整性的构建体。此外，原代肾脏祖细胞的包封导致高细胞活力，并随着时间的推移形成 3D 复杂结构。更重要的是，这种组织特异性基质还能够随着时间的推移影响细胞的生长和分化。总之，这些结果表明，未经修饰的 dKECM 生物墨水具有很大的潜力，可用于生物工程肾脏组织类似物，具有有前途的转化应用和/或模型系统。最终，这种策略可能会对生物医学领域产生更大的影响，用于开发使用其他组织的去细胞基质的生物工程替代品。
