The Persian Gulf Marine Biotechnology Research Center, The Persian Gulf Biomedical Sciences Research Institute, Bushehr University of Medical Sciences, Bushehr, Iran.
Percia Vista Biotechnology Company, Bushehr, Iran.
Cell Tissue Res. 2021 Nov;386(2):365-378. doi: 10.1007/s00441-021-03517-5. Epub 2021 Aug 23.
An automatic decellularization device was developed to perfuse and decellularize male rats' kidneys using both sodium lauryl ether sulfate (SLES) and sodium dodecyl sulfate (SDS) and to compare their efficacy in kidney decellularization and post-transplantation angiogenesis. Kidneys were perfused with either 1% SDS solution for 4 h or 1% SLES solution for 6 h. The decellularized scaffolds were stained with hematoxylin and eosin, periodic acid Schiff, Masson's trichrome, and Alcian blue to determine cell removal and glycogen, collagen, and glycosaminoglycan contents, respectively. Moreover, scanning electron microscopy was performed to evaluate the cell removal and preservation of microarchitecture of both SDS and SLES scaffolds. Additionally, DNA quantification assay was applied for all groups in order to measure residual DNA in the scaffolds and normal kidney. In order to demonstrate biocompatibility of the decellularized scaffolds, human umbilical cord mesenchymal stromal/stem cells (hUC-MSCs) were seeded on the scaffolds. In addition, the allotransplantation was performed in back muscle and angiogenesis was evaluated. Complete cell removal in both SLES and SDS groups was observed in scanning electron microscopy and DNA quantification assays. Moreover, the extracellular matrix (ECM) architecture of rat kidney in the SLES group was significantly preserved better than the SDS group. The hUC-MSCs were successfully migrated from the cell culture plate surface into the SDS and SLES decellularized scaffolds. The formation of blood vessels was observed in the kidney in both SLES and SDS decellularized kidneys. The better preservation of ECM than SDS introduces SLES as the solvent of choice for kidney decellularization.
开发了一种自动去细胞设备，用于使用十二烷基硫酸钠（SDS）和月桂基醚硫酸钠（SLES）灌注和去细胞化雄性大鼠的肾脏，并比较它们在肾脏去细胞化和移植后血管生成方面的效果。肾脏用 1% SDS 溶液灌注 4 小时或 1% SLES 溶液灌注 6 小时。用苏木精和伊红、过碘酸希夫、马松三色和阿利新蓝染色脱细胞支架，分别确定细胞去除和糖原、胶原和糖胺聚糖含量。此外，还进行了扫描电子显微镜检查，以评估 SDS 和 SLES 支架的细胞去除和微观结构保存情况。此外，对所有组应用 DNA 定量测定法，以测量支架和正常肾脏中残留的 DNA。为了证明脱细胞支架的生物相容性，将人脐带间充质基质/干细胞（hUC-MSCs）接种在支架上。此外，进行同种异体移植到背部肌肉，并评估血管生成。在扫描电子显微镜和 DNA 定量测定中，在 SLES 和 SDS 组中均观察到完全去除细胞。此外，SLES 组大鼠肾脏的细胞外基质（ECM）结构明显保存得更好。hUC-MSCs 成功地从细胞培养板表面迁移到 SDS 和 SLES 脱细胞支架中。在 SLES 和 SDS 脱细胞肾脏中均观察到肾脏中的血管形成。与 SDS 相比，ECM 的更好保存表明 SLES 是肾脏去细胞化的首选溶剂。
