Abolbashari Mehran, Agcaoili Sigrid M, Lee Mi-Kyung, Ko In Kap, Aboushwareb Tamer, Jackson John D, Yoo James J, Atala Anthony
Wake Forest Institute for Regenerative Medicine, Wake Forest School of Medicine, Medical Center Boulevard, Winston-Salem, NC 27157, USA.
Wake Forest Institute for Regenerative Medicine, Wake Forest School of Medicine, Medical Center Boulevard, Winston-Salem, NC 27157, USA; Molecular and Cellular Therapeutics Laboratory, Department of Urology, National Kidney and Transplantation Institute, Quezon City, Philippines.
Acta Biomater. 2016 Jan;29:52-61. doi: 10.1016/j.actbio.2015.11.026. Epub 2015 Nov 17.
The only definitive treatment for end stage renal disease is renal transplantation, however the current shortage of organ donors has resulted in a long list of patients awaiting transplant. Whole organ engineering based on decellularization/recellularization techniques has provided the possibility of creating engineered kidney constructs as an alternative to donor organ transplantation. Previous studies have demonstrated that small units of engineered kidney are able to maintain function in vivo. However, an engineered kidney with sufficient functional capacity to replace normal renal function has not yet been developed. One obstacle in the generation of such an organ is the development of effective cell seeding methods for robust colonization of engineered kidney scaffolds. We have developed cell culture methods that allow primary porcine renal cells to be efficiently expanded while maintaining normal renal phenotype. We have also established an effective cell seeding method for the repopulation of acellular porcine renal scaffolds. Histological and immunohistochemical analyses demonstrate that a majority of the expanded cells are proximal tubular cells, and the seeded cells formed tubule-like structures that express normal renal tubule phenotypic markers. Functional analysis revealed that cells within the kidney construct demonstrated normal renal functions such as re-adsorption of sodium and protein, hydrolase activity, and production of erythropoietin. These structural and functional outcomes suggest that engineered kidney scaffolds may offer an alternative to donor organ transplant.
Kidney transplantation is the only definitive treatment for end stage renal disease, however the current shortage of organ donors has limited the treatment. Whole organ engineering based on decellularization/recellularization techniques has provided the possibility of creating engineered kidney constructs as an alternative to donor organ transplantation. While previous studies have shown that small units of engineered kidneys are able to maintain function in animal studies, engineering of kidneys with sufficient functional capacity to replace normal renal function is still challenging due to inefficient cell seeding methods. This study aims to establish an effective cell seeding method using pig kidney cells for the repopulation of acellular porcine kidney scaffolds, suggesting that engineered kidneys may offer an alternative to donor organ transplant.
终末期肾病的唯一确定性治疗方法是肾移植，然而目前器官供体短缺导致大量患者等待移植。基于去细胞化/再细胞化技术的全器官工程为制造工程化肾脏构建体提供了可能性，以此作为供体器官移植的替代方案。先前的研究表明，工程化肾脏的小单元能够在体内维持功能。然而，尚未开发出具有足够功能能力以替代正常肾功能的工程化肾脏。生成这样一个器官的一个障碍是开发有效的细胞接种方法，以使工程化肾脏支架实现强大的细胞定植。我们已经开发出细胞培养方法，可使原代猪肾细胞在维持正常肾表型的同时高效扩增。我们还建立了一种有效的细胞接种方法，用于无细胞猪肾支架的再细胞化。组织学和免疫组织化学分析表明，大多数扩增细胞是近端肾小管细胞，接种的细胞形成了表达正常肾小管表型标志物的管状结构。功能分析显示，肾脏构建体内的细胞表现出正常的肾功能，如钠和蛋白质的重吸收、水解酶活性以及促红细胞生成素的产生。这些结构和功能结果表明，工程化肾脏支架可能为供体器官移植提供一种替代方案。
肾移植是终末期肾病的唯一确定性治疗方法，然而目前器官供体短缺限制了该治疗方法。基于去细胞化/再细胞化技术的全器官工程为制造工程化肾脏构建体提供了可能性，以此作为供体器官移植的替代方案。虽然先前的研究表明工程化肾脏的小单元在动物研究中能够维持功能，但由于细胞接种方法效率低下，构建具有足够功能能力以替代正常肾功能的肾脏仍然具有挑战性。本研究旨在建立一种使用猪肾细胞的有效细胞接种方法，用于无细胞猪肾支架的再细胞化，这表明工程化肾脏可能为供体器官移植提供一种替代方案。
