In Vivo Assessment of Laboratory-Grown Kidney Tissue Grafts.
In Vivo Assessment of Laboratory-Grown Kidney Tissue Grafts.
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DOI：
10.3390/bioengineering10111261
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发表时间：
2023-10-29
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干细胞的定向分化是产生用于再生疗法的肾组织的有吸引力的方法。目前，测试该组织再生潜力的最具信息性的平台是植入免疫受损啮齿动物的肾脏。干细胞衍生的肾组织在移植后血管化，但上皮小管之间的连接对于尿液从移植物传递到宿主收集系统至关重要，尚未得到证实。我们发现，小管融合的一个重要障碍是移植物和宿主之间界面处纤维状胶原的积累。作为一种筛选策略，以确定可以防止这种胶原蛋白积累的因素，我们建议将实验室生长的肾组织封装在纤维蛋白水凝胶中，并补充候选化合物，如重组蛋白，小分子，饲养细胞和基因治疗载体，以调节局部移植环境。我们证明了AAV-DJ血清型是一种有效的基因治疗载体的包膜下区域，它是特定的间质细胞在这个车厢。除了上皮小管融合的组织学评价外，我们还证明了两种尿生物标志物测定的特异性，其可用于检测近端肾单位（CD 59）和远端肾单位（尿调节蛋白）的人类特异性标志物，并且我们证明了人类移植物衍生的尿液沉积到小鼠收集系统中。使用本报告中描述的测试平台，将有可能系统地筛选具有促进移植物和宿主组织上皮融合的潜力的因子，并具有功能性活体读出。
Directed differentiation of stem cells is an attractive approach to generate kidney tissue for regenerative therapies. Currently, the most informative platform to test the regenerative potential of this tissue is engraftment into kidneys of immunocompromised rodents. Stem cell-derived kidney tissue is vascularized following engraftment, but the connection between epithelial tubules that is critical for urine to pass from the graft to the host collecting system has not yet been demonstrated. We show that one significant obstacle to tubule fusion is the accumulation of fibrillar collagens at the interface between the graft and the host. As a screening strategy to identify factors that can prevent this collagen accumulation, we propose encapsulating laboratory-grown kidney tissue in fibrin hydrogels supplemented with candidate compounds such as recombinant proteins, small molecules, feeder cells, and gene therapy vectors to condition the local graft environment. We demonstrate that the AAV-DJ serotype is an efficient gene therapy vector for the subcapsular region and that it is specific for interstitial cells in this compartment. In addition to the histological evaluation of epithelial tubule fusion, we demonstrate the specificity of two urine biomarker assays that can be used to detect human-specific markers of the proximal nephron (CD59) and the distal nephron (uromodulin), and we demonstrate the deposition of human graft-derived urine into the mouse collecting system. Using the testing platform described in this report, it will be possible to systematically screen factors for their potential to promote epithelial fusion of graft and host tissue with a functional intravital read-out.
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影响因子：
5.2
作者：
Bond KH;Chiba T;Wynne KPH;Vary CPH;Sims-Lucas S;Coburn JM;Oxburgh L
通讯作者：
Oxburgh L
影响因子：
13.2
作者：
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Zhuo, Xiaohui
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通讯作者：
Lipschutz, Joshua H.
影响因子：
5.4
作者：
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通讯作者：
Kay, Mark A.
影响因子：
1.9
作者：
Maung, Stephanie;El Sara, Ammar;Cukor, Daniel
通讯作者：
Cukor, Daniel