Department of Physiology, School of Medicine, Zahedan University of Medical Sciences, Zahedan, Iran; Department of Physiology, School of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran.
Department of Anatomy and Cell Biology, School of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran.
Life Sci. 2022 Apr 15;295:120167. doi: 10.1016/j.lfs.2021.120167. Epub 2021 Nov 22.
Regeneration of discarded human kidneys has been considered as an ideal approach to overcome organ shortage for the end-stage renal diseases (ESRDs). The aim of this study was to develop an effective method for preparation of kidney scaffolds that retain the matrix structure required for proliferation and importantly, differentiation of human adipose-derived mesenchymal stem cells (hAd-MSCs) into renal cells.
We first compared two different methods using triton X-100 and sodium dodecyl sulfate (SDS) for human kidney decellularization; followed by characterization of the prepared human renal extracellular matrix (ECM) scaffolds. Then, hAd-MSCs were seeded on the scaffolds and cultured for up to 3 weeks. Next, viability, proliferation, and migration of seeded hAd-MSCs underwent histological and scanning electron microscopy (SEM) assessments. Moreover, differentiation of hAd-MSCs into kidney-specific cell types was examined using immunohistochemistry (IHC) staining and qRT-PCR.
Our results indicated that triton X-100 was a more effective detergent for decellularization of human kidneys compared with SDS. Moreover, attachment and proliferation of hAd-MSCs within the recellularized human kidney scaffolds, were confirmed. Seeded cells expressed epithelial and endothelial differentiation markers, and qRT-PCR results indicated increased expression of platelet and endothelial cell adhesion molecule 1 (PECAM-1), paired box 2 (PAX2), and E-cadherine (E-CDH) as markers of differentiation into epithelial and endothelial cells.
These observations indicate the effectiveness of decellularization with triton X-100 to generate suitable human ECM renal scaffolds, which supported adhesion and proliferation of hAd-MSCs and could induce their differentiation towards a renal lineage.
将废弃的人类肾脏再生视为克服终末期肾脏疾病（ESRD）器官短缺的理想方法。本研究旨在开发一种有效的方法来制备保留用于增殖的基质结构的肾脏支架，重要的是，将人脂肪间充质干细胞（hAd-MSCs）分化为肾脏细胞。
我们首先比较了使用 Triton X-100 和十二烷基硫酸钠（SDS）两种不同方法对人肾脏脱细胞的效果；随后对制备的人肾脏细胞外基质（ECM）支架进行了表征。然后，将 hAd-MSCs 接种在支架上并培养长达 3 周。接下来，通过组织学和扫描电子显微镜（SEM）评估接种的 hAd-MSCs 的活力、增殖和迁移。此外，通过免疫组织化学（IHC）染色和 qRT-PCR 检查 hAd-MSCs 分化为肾脏特异性细胞类型的情况。
我们的结果表明，与 SDS 相比，Triton X-100 是一种更有效的人肾脏脱细胞去污剂。此外，在再细胞化的人肾脏支架内，证实了 hAd-MSCs 的附着和增殖。接种的细胞表达上皮和内皮分化标志物，qRT-PCR 结果表明，血小板和内皮细胞黏附分子 1（PECAM-1）、配对盒 2（PAX2）和 E-钙黏蛋白（E-CDH）的表达增加，这些标志物可分化为上皮和内皮细胞。
这些观察结果表明，使用 Triton X-100 进行脱细胞处理可有效产生合适的人 ECM 肾脏支架，支持 hAd-MSCs 的附着和增殖，并可诱导其向肾脏谱系分化。
