Center of Excellence in Translational Human Stem Cell Research, California National Primate Research Center, Davis, California 95616-8542, USA.
Tissue Eng Part A. 2010 Jul;16(7):2207-16. doi: 10.1089/ten.tea.2009.0602.
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.
本研究的目的是制备具有结构、力学和生理特性的脱细胞肾脏支架,这些特性是体外工程基本肾脏结构所必需的。采用 1%(v/v)十二烷基硫酸钠或 Triton X-100 处理胎儿、婴儿、青少年和成年恒河猴肾脏切片,进行定量和定性分析。脱细胞试剂和孵育温度的比较表明,4°C 的十二烷基硫酸钠在保持固有结构方面最有效。苏木精和伊红染色证实了细胞物质的去除,免疫组织化学显示了细胞外基质蛋白的固有表达模式的保留,包括硫酸乙酰肝素蛋白聚糖、纤连蛋白、胶原 I 和 IV 以及层粘连蛋白。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降低。与年龄匹配的脱细胞肾脏支架一起分层培养胎儿肾脏外植体,证明支架具有支持 Pax2+/vimentin+细胞附着和迁移以重新填充支架的能力。这些发现表明,脱细胞肾脏切片保留了作为三维支架使用所必需的关键结构和功能特性,并促进了细胞再定植。此外,本研究为肾脏组织工程和修复的再生医学策略的发展提供了初步步骤。