Decellularized Rhesus Monkey Kidney as a Three-Dimensional Scaffold for Renal Tissue Engineering
Decellularized Rhesus Monkey Kidney as a Three-Dimensional Scaffold for Renal Tissue Engineering
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DOI：
10.1089/ten.tea.2009.0602
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发表时间：
2010-07-01
影响因子：
4.1
通讯作者：
Tarantal, Alice F.
中科院分区：
文献类型：
--
作者：
Nakayama, Karina H.;Batchelder, Cynthia A.;Tarantal, Alice F.
本研究的目的是制备一种脱细胞肾支架，该支架具有结构、机械和生理特性，是体外工程基本肾脏结构所必需的。用1% （v/v）十二烷基硫酸钠或Triton X-100处理胎儿、婴儿、幼猴和成年恒河猴肾脏切片，进行定量和定性分析。脱细胞剂和孵育温度的比较表明，在4℃下十二烷基硫酸钠对保存天然结构最有效。苏木精和伊红染色证实了细胞物质的去除，免疫组织化学证实了细胞外基质蛋白的天然表达模式的保存，包括硫酸肝素蛋白聚糖、纤维连接蛋白、I型和IV型胶原蛋白和层粘连蛋白。生物力学测试显示，与新鲜肾脏相比，脱细胞肾脏的压缩模量有所下降。在年龄匹配的脱细胞肾支架上分层胎儿肾移植，证明了支架支持Pax2+/ vimentin+细胞附着和迁移以使支架再细胞化的能力。这些发现表明，去细胞肾切片保留了作为三维支架和促进细胞再生所必需的关键结构和功能特性。此外，本研究为开发新的肾组织工程和修复再生医学策略提供了初步的步骤。
The goal of this study was the production of a decellularized kidney scaffold with structural, mechanical, and physiological properties necessary for engineering basic renal structures in vitro. Fetal, infant, juvenile, and adult rhesus monkey kidney sections were treated with either 1% (v/v) sodium dodecyl sulfate or Triton X-100 followed by quantitative and qualitative analysis. Comparison of decellularization agents and incubation temperatures demonstrated sodium dodecyl sulfate at 4 degrees C to be most effective in preserving the native architecture. Hematoxylin and eosin staining confirmed the removal of cellular material, and immunohistochemistry demonstrated preservation of native expression patterns of extracellular matrix proteins, including heparan sulfate proteoglycan, fibronectin, collagen types I and IV, and laminin. Biomechanical testing revealed a decrease in the compressive modulus of decellularized compared to fresh kidneys. Layering of fetal kidney explants on age-matched decellularized kidney scaffolds demonstrated the capacity of the scaffold to support Pax2+/ vimentin+ cell attachment and migration to recellularize the scaffold. These findings demonstrate that decellularized kidney sections retain critical structural and functional properties necessary for use as a three-dimensional scaffold and promote cellular repopulation. Further, this study provides the initial steps in developing new regenerative medicine strategies for renal tissue engineering and repair.