Clinical-Grade Isolated Human Kidney Perivascular Stromal Cells as an Organotypic Cell Source for Kidney Regenerative Medicine

Abstract

Mesenchymal stromal cells (MSCs) are immunomodulatory and tissue homeostatic cells that have shown beneficial effects in kidney diseases and transplantation. Perivascular stromal cells (PSCs) identified within several different organs share characteristics of bone marrow‐derived MSCs (BM‐MSCs). These PSCs may also possess tissue‐specific properties and play a role in local tissue homeostasis. We hypothesized that human kidney‐derived PSCs (hkPSCs) would elicit improved kidney repair in comparison with BM‐MSCs. Here we introduce a novel, clinical‐grade isolation method of hkPSCs from cadaveric kidneys by enriching for the perivascular marker, NG2. hkPSCs show strong transcriptional similarities to BM‐MSCs but also show organotypic expression signatures, including the HoxD10 and HoxD11 nephrogenic transcription factors. Comparable to BM‐MSCs, hkPSCs showed immunosuppressive potential and, when cocultured with endothelial cells, vascular plexus formation was supported, which was specifically in the hkPSCs accompanied by an increased NG2 expression. hkPSCs did not undergo myofibroblast transformation after exposure to transforming growth factor‐β, further corroborating their potential regulatory role in tissue homeostasis. This was further supported by the observation that hkPSCs induced accelerated repair in a tubular epithelial wound scratch assay, which was mediated through hepatocyte growth factor release. In vivo, in a neonatal kidney injection model, hkPSCs reintegrated and survived in the interstitial compartment, whereas BM‐MSCs did not show this potential. Moreover, hkPSCs gave protection against the development of acute kidney injury in vivo in a model of rhabdomyolysis‐mediated nephrotoxicity. Overall, this suggests a superior therapeutic potential for the use of hkPSCs and their secretome in the treatment of kidney diseases. Stem Cells Translational Medicine 2017;6:405–418

Concise Review: The Endothelial Cell Extracellular Matrix Regulates Tissue Homeostasis and Repair

“…The composition of this latter layer is primarily controlled by resident tissue fibroblasts, that act as a signaling hub for tissue homeostasis and repair. Like the endothelium, they have an organ‐specific phenotype where anatomical localization is related to the specific transcriptome profiles, such as embryonic patterning genes, as well as function . While these cells intimately communicate with tissue cells and the immune system to preserve an anti‐inflammatory and trophic milieu, they can respond to injury by differentiating into proliferating myofibroblasts and eventually converting into a nonproliferating, matrix‐producing phenotype.…”

Lowering the increased intracellular pH of human-induced pluripotent stem cell-derived endothelial cells induces formation of mature Weibel-Palade bodies

“…Human kidney-derived perivascular stromal cells (hkPSCs) were isolated from transplant-grade kidneys discarded for as surgical waste. 35 fluorescence-activated cell sorting (FACS) confirmed that homogeneous NG2-positive hkPSC populations between passages 4 and 8 and hMVEC between passages 6 and 8 were cultured in EC-SFM full medium at 37 C and 5% CO 2 as described in detail previously. 34…”

“…RNA isolation was achieved using an RNeasy mini kit (Qiagen, Hilden, Germany), and quantitative polymerase chain reaction (qPCR) was performed as previously described. 35 Forward and reversed VWF, KLF2, and MCT1 primer sequences are depicted in Table 1. Ct values were normalized by the Ct of GAPDH.…”

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

“…This enhanced wound repair was mediated through hepatocyte growth factor release. Moreover, kPSCs ameliorated kidney injury in a mouse model of acute kidney injury 15 . Therefore, kPSCs seem to have superior renal repair capacities and are an interesting new source for cell therapy in kidney diseases.…”

“…Perivascular stromal cells can also be isolated from the mouse 12 13 14 and human kidney 15 16 . We extensively characterized kPSCs and compared these to bmMSCs.…”