Uzarski Joseph S, Beck Emily C, Russell Emily E, Vanderslice Ethan J, Holzner Matthew L, Wadhera Vikram, Adamson Dylan, Shapiro Ron, Davidow Dominique S, Ross Jeff J, Florman Sander S
Miromatrix Medical Inc., Eden Prairie, MN, United States.
Icahn School of Medicine at Mount Sinai, Recanati/Miller Transplantation Institute, New York, NY, United States.
Front Bioeng Biotechnol. 2023 Jun 14;11:1184408. doi: 10.3389/fbioe.2023.1184408. eCollection 2023.
Despite progress in whole-organ decellularization and recellularization, maintaining long-term perfusion remains a hurdle to realizing clinical translation of bioengineered kidney grafts. The objectives for the present study were to define a threshold glucose consumption rate (GCR) that could be used to predict graft hemocompatibility and utilize this threshold to assess the performance of clinically relevant decellularized porcine kidney grafts recellularized with human umbilical vein endothelial cells (HUVECs). Twenty-two porcine kidneys were decellularized and 19 were re-endothelialized using HUVECs. Functional revascularization of control decellularized ( = 3) and re-endothelialized porcine kidneys ( = 16) was tested using an porcine blood flow model to define an appropriate metabolic glucose consumption rate (GCR) threshold above which would sustain patent blood flow. Re-endothelialized grafts ( = 9) were then transplanted into immunosuppressed pigs with perfusion measured using angiography post-implant and on days 3 and 7 with 3 native kidneys used as controls. Patent recellularized kidney grafts underwent histological analysis following explant. The glucose consumption rate of recellularized kidney grafts reached a peak of 39.9 ± 9.7 mg/h at 21 ± 5 days, at which point the grafts were determined to have sufficient histological vascular coverage with endothelial cells. Based on these results, a minimum glucose consumption rate threshold of 20 mg/h was set. The revascularized kidneys had a mean perfusion percentage of 87.7% ± 10.3%, 80.9% ± 33.1%, and 68.5% ± 38.6% post-reperfusion on Days 0, 3 and 7, respectively. The 3 native kidneys had a mean post-perfusion percentage of 98.4% ± 1.6%. These results were not statistically significant. This study is the first to demonstrate that human-scale bioengineered porcine kidney grafts developed via perfusion decellularization and subsequent re-endothelialization using HUVEC can maintain patency with consistent blood flow for up to 7 days . These results lay the foundation for future research to produce human-scale recellularized kidney grafts for transplantation.
尽管在全器官去细胞化和再细胞化方面取得了进展，但维持长期灌注仍然是生物工程肾移植实现临床转化的一个障碍。本研究的目的是确定一个可用于预测移植物血液相容性的葡萄糖消耗率（GCR）阈值，并利用该阈值评估用人脐静脉内皮细胞（HUVEC）再细胞化的临床相关去细胞化猪肾移植物的性能。将22个猪肾去细胞化，其中19个用HUVEC进行再内皮化。使用猪血流模型测试对照去细胞化猪肾（n = 3）和再内皮化猪肾（n = 16）的功能性血管重建，以确定一个适当的代谢葡萄糖消耗率（GCR）阈值，高于该阈值可维持通畅的血流。然后将再内皮化移植物（n = 9）移植到免疫抑制的猪体内，植入后使用血管造影术测量灌注情况，并在第3天和第7天进行测量，将3个天然肾用作对照。再细胞化肾移植物在取出后进行组织学分析。再细胞化肾移植物的葡萄糖消耗率在21±5天时达到峰值39.9±9.7mg/h，此时确定移植物具有足够的内皮细胞组织学血管覆盖。基于这些结果，设定了20mg/h的最低葡萄糖消耗率阈值。再血管化肾在再灌注后第0天、第3天和第7天的平均灌注百分比分别为87.7%±10.3%、80.9%±33.1%和68.5%±38.6%。3个天然肾的平均灌注后百分比为98.4%±1.6%。这些结果无统计学意义。本研究首次证明，通过灌注去细胞化和随后使用HUVEC进行再内皮化开发的人体规模生物工程猪肾移植物可维持通畅的血流长达7天。这些结果为未来生产用于移植的人体规模再细胞化肾移植物的研究奠定了基础。
