Chang William Gee, Fornoni Alessia, Tietjen Gregory, Mendez Julio J, Niklason Laura E, Saltzman W Mark, Pober Jordan S
1 Department of Medicine and Section of Nephrology, Yale University School of Medicine , New Haven, Connecticut.
2 Division of Nephrology and Hypertension and Diabetes Research Institute, University of Miami Miller School of Medicine , Miami, Florida.
Tissue Eng Part A. 2015 Nov;21(21-22):2673-9. doi: 10.1089/ten.TEA.2015.0060. Epub 2015 Oct 1.
Kidney transplantation is often the most effective therapy for end-stage renal disease, but there are not enough donor organs to meet the rising demand. Tissue engineering of kidneys is a potential solution to this organ shortage. Achieving microvascular perfusion has been a major barrier to engineering tissues beyond thin muscularized sheets such as the bladder wall. Our laboratory has previously reported that human umbilical vein endothelial cells (ECs) transduced with the antiapoptotic protein Bcl-2 will spontaneously organize into perfused microvessels within type I collagen gels when implanted in immunodeficient mice. To test if this system can be used to perfuse more complex structures, we combined Bcl-2-transduced ECs (Bcl-2-ECs) with renal glomeruli, the specialized vascular filtration units of the kidney. Microdissected green fluorescent protein-expressing rat glomeruli suspended in type I collagen gels were implanted within immunodeficient mice with or without the inclusion of Bcl-2-ECs. Survival of rat glomeruli was enhanced by coimplantation with Bcl-2-ECs. Intravital rhodamine dextran injections demonstrated that surviving glomeruli were perfused through Bcl-2-EC-derived microvessels. Perfused glomeruli maintained podocin staining, but transmission electron microscopy revealed endothelial swelling and podocyte foot process effacement. Anastomosis of microvessels derived from Bcl-2-ECs with glomerular capillaries provides proof of concept that self-assembled microvessels can perfuse specialized organ structures such as glomeruli, but that perfusion alone may be insufficient to maintain normal structure.
肾移植通常是终末期肾病最有效的治疗方法,但供体器官不足,无法满足不断增长的需求。肾脏组织工程是解决这种器官短缺问题的一个潜在办法。实现微血管灌注一直是构建除膀胱壁等薄肌化组织之外的其他组织的主要障碍。我们实验室之前报道过,转导了抗凋亡蛋白Bcl-2的人脐静脉内皮细胞(ECs)植入免疫缺陷小鼠体内后,会在I型胶原凝胶中自发组织形成有灌注功能的微血管。为了测试该系统是否可用于灌注更复杂的结构,我们将转导了Bcl-2的内皮细胞(Bcl-2-ECs)与肾小球(肾脏的特殊血管滤过单位)相结合。将悬浮于I型胶原凝胶中的经显微切割的绿色荧光蛋白表达大鼠肾小球植入免疫缺陷小鼠体内,植入时或包含或不包含Bcl-2-ECs。与Bcl-2-ECs共同植入可提高大鼠肾小球的存活率。活体注射罗丹明葡聚糖显示,存活的肾小球通过源自Bcl-2-ECs的微血管实现了灌注。灌注的肾小球保持足突蛋白染色,但透射电子显微镜显示内皮肿胀和足细胞足突消失。源自Bcl-2-ECs的微血管与肾小球毛细血管吻合,证明了自组装微血管可灌注诸如肾小球等特殊器官结构的概念,但仅灌注可能不足以维持正常结构。