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一种用于人肾周血管基质细胞的新型临床级分离方法。

A Novel Clinical Grade Isolation Method for Human Kidney Perivascular Stromal Cells.

作者信息

Leuning Daniëlle G, Lievers Ellen, Reinders Marlies E J, van Kooten Cees, Engelse Marten A, Rabelink Ton J

机构信息

Department of Internal Medicine, Leiden University Medical Centre.

Department of Internal Medicine, Leiden University Medical Centre;

出版信息

J Vis Exp. 2017 Aug 7(126):55841. doi: 10.3791/55841.

Abstract

Mesenchymal Stromal Cells (MSCs) are tissue homeostatic and immune modulatory cells that have shown beneficial effects in kidney diseases and transplantation. Perivascular Stromal Cells (PSCs) share characteristics with bone marrow MSCs (bmMSCs). However, they also possess, most likely due to local imprinting, tissue-specific properties and play a role in local tissue homeostasis. This tissue specificity may result in tissue specific repair, also within the human kidney. We previously showed that human kidney PSCs (kPSCs) have enhanced kidney epithelial wound healing whereas bmMSCs did not have this potential. Moreover, kPSCs can ameliorate kidney injury in vivo. Therefore, kPSCs constitute an interesting source for cell therapy, particularly for kidney diseases and renal transplantation. Here we show the detailed isolation and culture method for kPSCs from transplant-grade human kidneys based on whole-organ perfusion of digestive enzymes via the renal artery and enrichment for the perivascular marker NG2. In this way, large cell quantities can be obtained that are suitable for cellular therapy.

摘要

间充质基质细胞(MSCs)是组织稳态和免疫调节细胞,已在肾脏疾病和移植中显示出有益作用。血管周围基质细胞(PSCs)与骨髓间充质干细胞(bmMSCs)具有共同特征。然而,它们很可能由于局部印记而具有组织特异性特性,并在局部组织稳态中发挥作用。这种组织特异性可能导致组织特异性修复,在人类肾脏中也是如此。我们之前表明,人肾PSCs(kPSCs)具有增强的肾上皮伤口愈合能力,而bmMSCs则没有这种潜力。此外,kPSCs可以在体内改善肾损伤。因此,kPSCs构成了一种有趣的细胞治疗来源,特别是对于肾脏疾病和肾移植。在这里,我们展示了从移植级人肾中分离和培养kPSCs的详细方法,该方法基于通过肾动脉对消化酶进行全器官灌注并富集血管周围标志物NG2。通过这种方式,可以获得大量适合细胞治疗的细胞。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b00b/5614134/e0b4b4d956e8/jove-126-55841-0.jpg

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