Iwai S, Sakonju I, Okano S, Teratani T, Kasahara N, Yokote S, Yokoo T, Kobayash E
Laboratory of Small Animal Surgery I, School of Veterinary Medicine, Kitasato University, Towada, Aomori, Japan.
Laboratory of Small Animal Surgery I, School of Veterinary Medicine, Kitasato University, Towada, Aomori, Japan.
Transplant Proc. 2014 Jun;46(5):1578-84. doi: 10.1016/j.transproceed.2013.12.068.
Mesenchymal stem cells (MSCs) have been applied to the treatment of various diseases, and MSC administration in marginal donor grafts may help avoid the ischemia-reperfusion injury associated with solid organ transplants. Given the reports of side effects after intravenous MSC administration, local MSC administration to the target organ might be a better approach. We administered adipose tissue-derived MSCs (AT-MSCs) ex vivo to donor rat kidneys obtained after cardiac death (CD).
Using male Lewis rats (8-10 weeks), and a marginal transplant model of 1hr CD plus 1hr sub-normothermic ET-Kyoto solution preservation were conducted. AT-MSCs obtained from double-reporter (luciferase-LacZ) transgenic Lewis rats were injected either systemically (1.0 × 10(6) cells/0.5 mL) to bilaterally nephrectomized recipient rats that had received a marginal kidney graft (n = 6), or locally via the renal artery (500 μL ET-Kyoto solution containing the same number of AT-MSCs) to marginal kidney grafts, which were then preserved (1 hour; 22°C) before being transplanted into bilaterally nephrectomized recipient rats (n = 8). Serum was collected to assess the therapeutic effects of AT-MSC administration, and the recipients of rats surviving to Day 14 were separately evaluated histopathologically. Follow-up was by in vivo imaging and histological LacZ staining, and tumor formation was evaluated in MSC-injected rats at 3 months.
Systemic injection of MSC did not improve recipient survival. In vivo imaging showed MSCs trapped in the lung that later became undetectable. Ex vivo injection of MSCs did show a benefit without adverse effects. At Day 14 after RTx, 75% of the rats in the AT-MSC-injected group (MSC[+]) had survived, whereas 50% of the rats in the AT-MSC-non-injected group (MSC[-]) had died. Renal function in the MSC(+) group was improved compared with that in the MSC(-) group at Day 4. LacZ staining revealed AT-MSCs attached to the renal tubules at 24 hours after RTx that later became undetectable. Histopathologic examination showed little difference in fibrosis between the groups at Day 14. No teratomas or other abnormalities were seen at 3 months.
间充质干细胞（MSCs）已被应用于多种疾病的治疗，在边缘供体移植物中施用MSCs可能有助于避免与实体器官移植相关的缺血再灌注损伤。鉴于静脉注射MSCs后出现副作用的报道，将MSCs局部施用于靶器官可能是一种更好的方法。我们将体外培养的脂肪组织来源的间充质干细胞（AT-MSCs）施用于心脏死亡（CD）后获取的供体大鼠肾脏。
使用雄性Lewis大鼠（8-10周龄），进行1小时心脏死亡加1小时亚常温ET-京都溶液保存的边缘移植模型。从双报告基因（荧光素酶-LacZ）转基因Lewis大鼠获得的AT-MSCs，要么全身注射（1.0×10(6)个细胞/0.5 mL）给接受边缘肾移植的双侧肾切除受体大鼠（n = 6），要么通过肾动脉局部注射（500 μL含相同数量AT-MSCs的ET-京都溶液）给边缘肾移植物，然后在移植到双侧肾切除受体大鼠（n = 8）之前保存（1小时；22°C）。收集血清以评估AT-MSC给药的治疗效果，对存活至第14天的大鼠受体分别进行组织病理学评估。通过体内成像和组织学LacZ染色进行随访，并在3个月时评估注射MSCs大鼠的肿瘤形成情况。
全身注射MSCs并未提高受体存活率。体内成像显示MSCs被困在肺中，随后无法检测到。体外注射MSCs确实显示出有益效果且无不良反应。肾移植后第14天，注射AT-MSCs组（MSC[+]）的大鼠中有75%存活，而未注射AT-MSCs组（MSC[-]）的大鼠中有50%死亡。肾移植后第4天，MSC(+)组的肾功能比MSC(-)组有所改善。LacZ染色显示肾移植后24小时AT-MSCs附着于肾小管，随后无法检测到。组织病理学检查显示第14天时两组间纤维化差异不大。3个月时未观察到畸胎瘤或其他异常。
