人胚胎干细胞源内皮细胞通过刺激肾祖细胞增殖保护肾脏急性缺血

贾晓花; 吕赫; 李辰; 冯国伟; 姚欣鹏; 毛丽娜; 柯亭羽; 车永哲; 徐勇; 李宗金; 孔德领

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Human Embryonic Stem Cells-Derived Endothelial Cell Therapy Facilitates Kidney Regeneration by Stimulating Resident Stem Cell Proliferation in Acute Kidney Injury

首发时间：2012-12-27

Jia Xiaohua ¹
JIA Xiaohua(1980.10.5)
Lv He ² Li Chen ³ Feng Guowei ⁴ Yao Xinpeng ¹ Mao Lina ¹ Ke Tingyu ¹ Che Yongzhe ¹ Xv Yong ⁴ Li Zongjin ¹
LI Zongjin
Kong Deling ¹
KONG Deling

1、the Key Laboratory of Bioactive Materials, Ministry of Education, Nankai University, Tianjin 300071
2、College of Basic Medicine, Yanbian University, Yanji, Jilin 133000
3、Diabetes Research Group, King's College London, UK
4、Department of Urology, Second Hospital of Tianjin Medical University, Tianjin Institute of Urology, Tianjin 300071

Abstract：Endothelial cell therapy has been implicated to enhance tissue regeneration and vascularization in ischemic kidney. However, no published study has yet examined direct effects of endothelial cell treatment in kidney recovery. This study investigated the therapeutic efficacy of endothelial cells in a mouse model with acute kidney injury (AKI). Thus, human embryonic stem cells-derived endothelial cells (hESC-ECs) labeled with a reporter system encoding a double fusion reporter gene for firefly luciferase (Fluc) and green fluorescent protein (GFP) were characterized by Fluc imaging and immunofluoresence staining. Cultured hESC-ECs (1×106) were injected into ischemic kidney shortly after AKI. Survival of the transplanted hESC-ECs was monitored in vivo from day 1 to 14 after endothelial cell transplantation and potential impact of hESC-EC treatment on renal regeneration was assessed by histological analyses. We report that a substantial level of bioluminescence activity was detected 24 hours after hESC-EC injection followed by a gradual decline from day 1 to 14. Human ESC-ECs markedly accelerated kidney cell proliferation in response to ischaemia-induced damage, indicated by an elevated number of BrdU+ cells. Co-expression of Sca-1, a kidney stem cell proliferation marker, and BrdU further suggested that the observed stimulation in renal cell regeneration was, at least in part, due to increased proliferation of renal resident stem cells especially within the medullary cords and arteriole. Differentiation of hESC-ECs to smooth muscle cells was also observed at an early stage of kidney recovery. In summary, our results suggest that endothelial cell therapy facilitates kidney recovery by promoting vascularization, trans-differentiation and endogenous renal stem cell proliferation in AKI.

keywords： Molecular imaging Kidney ischemia Renal stem cells Endothelial cells Cell transplantation

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人胚胎干细胞源内皮细胞通过刺激肾祖细胞增殖保护肾脏急性缺血

贾晓花 ¹
JIA Xiaohua(1980.10.5)
吕赫 ² 李辰 ³ 冯国伟 ⁴ 姚欣鹏 ¹ 毛丽娜 ¹ 柯亭羽 ¹ 车永哲 ¹ 徐勇 ⁴ 李宗金 ¹
LI Zongjin
孔德领 ¹
KONG Deling

1、生物活性材料教育部重点实验室,生命科学学院,南开大学,天津 300071
2、基础医学院,延边大学,延吉,吉林 133000
3、糖尿病研究组,国王学院,伦敦,英国
4、泌尿外科,天津医科大学附属第二医院,天津泌尿研究所,天津,300071

摘要：内皮细胞治疗可改善血管再生，尤其是促进功能性微脉管系统的再生，被认为是一个很有潜力的治疗方案。然而，还未有研究直接检查内皮细胞治疗对肾复苏的影响作用。本论文主要探讨内皮细胞在小鼠急性肾损伤模型中的疗效。首先将人胚胎干细胞中提取的内皮细胞(hESC-ECs)用一个荧光素酶和绿色荧光蛋白(GFP)共同编码的双融合报告基因标记。培养的hESC-ECs(1×106)被注射到急性缺血/再灌注肾，通过活体成像技术监控体内hESC-ECs从移植后的第1天到第14天的生存情况。通过组织学分析评估内皮细胞移植对hESC-EC治疗肾再生潜在的影响。我们发现hESC-EC注入24小时内可见稳定的生物荧光活动，但随着时间推移荧光信号逐渐减弱。通过计数BrdU+ 细胞数发现hESC-ECs显著加速缺血损伤区的肾细胞增殖。Sca-1(肾脏干细胞标记)和BrdU的共表达染色发现hESC-EC治疗肾再生潜在的影响部分要归功于增加肾干细胞的增殖。hESC-ECs在注射入动物体内的早期阶段可分化平滑肌细胞。总之，我们的结果表明，内皮细胞疗法通过促进血管化、转分化和内生肾干细胞的增殖促进肾功能恢复。

关键词：分子成像肾缺血肾干细胞内皮细胞细胞移植

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1. Doctoral Fund of Ministry of Education of China （20090031110024）

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Jia Xiaohua,Lv He,Li Chen,et al. Human Embryonic Stem Cells-Derived Endothelial Cell Therapy Facilitates Kidney Regeneration by Stimulating Resident Stem Cell Proliferation in Acute Kidney Injury[EB/OL]. Beijing:Sciencepaper Online[2012-12-27]. https://www.paper.edu.cn/releasepaper/content/201212-926.

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论文编号	201212-926
论文题目	人胚胎干细胞源内皮细胞通过刺激肾祖细胞增殖保护肾脏急性缺血
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