Choi Minji, Yang Yu-Bin, Park Seongsu, Rahaman Sohanur, Tripathi Garima, Lee Byong-Taek
Department of Regenerative Medicine, College of Medicine, Soonchunhyang University, South Korea.
Institute of Tissue Regeneration, Soonchunhyang University, Cheonan, South Korea.
Mater Today Bio. 2022 Oct 18;17:100464. doi: 10.1016/j.mtbio.2022.100464. eCollection 2022 Dec 15.
In recent era, many researches on implantable bio-artificial organs has been increased owing to large gap between donors and receivers. Comprehensive organ based researches on perfusion culture for cell injury using different flow rate have not been conducted at the cellular level. The present study investigated the co-culture of rat glomerulus endothelial cell (rGEC) and rat bone marrow mesenchymal stem cells (rBMSC) to develop micro vascularization in the kidney scaffolds culturing by bioreactor system. To obtain kidney scaffold, extracted rat kidneys were decellularized by 1% sodium dodecyl sulfate (SDS), 1% triton X-100, and distilled water. Expanded rGECs were injected through decellularized kidney scaffold artery and cultured using bioreactor system. Vascular endothelial cells adhered and proliferated on the renal ECM scaffold in the bioreactor system for 3, 7 and 14 days. Static, 1 ml/min and 2 ml/min flow rates (FR) were tested and among them, 1 ml/min flow rate was selected based on cell viability, glomerulus character, inflammation/endothelialization proteins expression level. However, the flow injury was still existed on primary cell cultured at vessel in kidney scaffold. Therefore, co-culture of rGEC + rBMSC found suitable to possibly solve this problem and resulted increased cell proliferation and micro-vascularization in the glomerulus, reducing inflammation and cell death which induced by flow injury. The optimized perfusion rate under rGEC + rBMSC co-culture conditions resulted in enhanced endocellularization to make ECM derived implantable renal scaffold and might be useful as a way of treatment of the acute renal failure.
近年来,由于供体与受体之间存在巨大差距,关于可植入生物人工器官的研究不断增加。尚未在细胞水平上针对使用不同流速进行细胞损伤的灌注培养开展基于综合器官的研究。本研究调查了大鼠肾小球内皮细胞(rGEC)和大鼠骨髓间充质干细胞(rBMSC)的共培养,以在通过生物反应器系统培养的肾支架中实现微血管化。为了获得肾支架,提取的大鼠肾脏用1%十二烷基硫酸钠(SDS)、1% Triton X-100和蒸馏水进行脱细胞处理。将扩增的rGEC通过脱细胞肾支架动脉注入,并使用生物反应器系统进行培养。血管内皮细胞在生物反应器系统中的肾ECM支架上附着并增殖3、7和14天。测试了静态、1毫升/分钟和2毫升/分钟的流速(FR),其中基于细胞活力、肾小球特征、炎症/内皮化蛋白表达水平选择了1毫升/分钟的流速。然而,在肾支架血管中培养的原代细胞上仍存在流动损伤。因此,rGEC + rBMSC的共培养被发现可能适合解决这个问题,并导致肾小球中细胞增殖增加和微血管化,减少由流动损伤引起的炎症和细胞死亡。rGEC + rBMSC共培养条件下的优化灌注率导致细胞内形成增强,从而制成ECM衍生的可植入肾支架,可能作为治疗急性肾衰竭的一种方法。