Department of Stem Cells & Regenerative Medicine, D Y Patil Education Society (Deemed to be University), Kolhapur, Maharashtra, India.
Department of Radiology, D Y Patil Medical College, D Y Patil Education Society (Deemed to be University), Kolhapur, Maharashtra, India.
Adv Exp Med Biol. 2024;1450:77-92. doi: 10.1007/5584_2023_785.
A significant hurdle for kidney tissue engineering is reproducing the complex three-dimensional structure of the kidney. In our study, a stepwise approach of generating a reproducible Xeno kidney scaffold from a goat kidney is described, which can be implanted and recellularized by host cells. We have proposed a combination of sodium dodecyl sulfate and Triton-X-100-based protocol to generate a reproducible Xeno kidney scaffold, which was then analyzed by histology, DNA quantification, SEM, and renal angiography. Further, a small portion from the cortico-medullar region of the acellular scaffold was implanted in the rat's kidney subcapsular pocket for a period of 1 month, to check the recruitment of host cells into the scaffold. Post implantation, the extracellular matrix of the scaffold was well preserved and it did not induce any damage or inflammation in the native kidney. Implantation of the Xeno scaffold resulted in apparent early vascularization which helped in the recruitment of the host cells, which was characterized by histology, immunohistochemistry, and scanning electron microscopy. Implanted Xeno scaffold showed AQP-1, Nephrin, α-SMA, and VEGF expression in proximal tubules and renal glomerulus. Importantly, Ki-67 and WTAP-expressing cells were also observed near proximal tubules suggesting a high level of proliferation in the scaffold. Thus, showing the potential of Xeno kidney development that can be recellularized by the host cell to engineer into a functional kidney.
肾组织工程的一个重大障碍是复制肾脏的复杂三维结构。在我们的研究中,描述了一种从山羊肾脏生成可重复的异种肾脏支架的逐步方法,该支架可以被宿主细胞植入和再细胞化。我们提出了一种基于十二烷基硫酸钠和 Triton-X-100 的组合方案来生成可重复的异种肾脏支架,然后通过组织学、DNA 定量、SEM 和肾脏血管造影对其进行分析。此外,将脱细胞支架的皮质-髓质区域的一小部分植入大鼠肾脏的肾包膜下袋中 1 个月,以检查宿主细胞是否进入支架。植入后,支架的细胞外基质得到了很好的保存,并且没有在原生肾脏中引起任何损伤或炎症。异种支架的植入导致明显的早期血管化,有助于宿主细胞的募集,这一过程通过组织学、免疫组织化学和扫描电子显微镜得到了特征化。植入的异种支架在近端肾小管和肾小球中显示出 AQP-1、Nephrin、α-SMA 和 VEGF 的表达。重要的是,还在近端肾小管附近观察到 Ki-67 和 WTAP 表达的细胞,表明支架中有高水平的增殖。因此,表明异种肾脏发展的潜力,即可以通过宿主细胞再细胞化来构建成功能性肾脏。