Abstract
Mesenchymal stromal/stem cells (MSCs) are immature multipotent cells, which represent a rare population in the perivascular niche within nearly all tissues. The most abundant source to isolate MSCs is adipose tissue. Currently, perirenal adipose tissue is rarely described as the source of MSCs. MSCs were isolated from perirenal adipose tissue (prASCs) from patients undergoing tumor nephrectomies, cultured and characterized by flow cytometry and their differentiation potential into adipocytes, chondrocytes, osteoblasts and epithelial cells. Furthermore, prASCs were stimulated with lipopolysaccharide (LPS), lipoteichoic acid (LTA) or a mixture of cytokines (cytomix). In addition, prASC susceptibility to human cytomegalovirus (HCMV) was investigated. The expression of inflammatory readouts was estimated by qPCR and immunoassay. HCMV infection was analyzed by qPCR and immunostaining. Characterization of cultured prASCs shows the cells meet the criteria of MSCs and prASCs can undergo trilineage differentiation. Cultured prASCs can be induced to differentiate into epithelial cells, shown by cytokeratin 18 expression. Stimulation of prASCs with LPS or cytomix suggests the cells are capable of initiating an inflammation-like response upon stimulation with LPS or cytokines, whereas, LTA did not induce a significant effect on the readouts (ICAM-1, IL-6, TNFα, MCP-1 mRNA and IL-6 protein). HCMV broadly infects prASCs, showing a viral load dependent cytopathological effect (CPE). Our current study summarizes the isolation and culture of prASCs, clearly characterizes the cells, and demonstrates their immunomodulatory potential and high permissiveness for HCMV.
Highlights
Mesenchymal stromal/stem cells (MSCs) are immature multipotent stromal cells, which represent a rare population in the perivascular niche within fully specialized tissues throughout theCells 2019, 8, 1346; doi:10.3390/cells8111346 www.mdpi.com/journal/cellsCells 2019, 8, 1346 whole body [1]
The present study further investigated the immunomodulatory potential of perirenal adiposederived stromal/stem cells (prASCs) after stimulation with lipopolysaccharide (LPS), lipoteichoic acid (LTA), a mixture of cytokines, or infection with
We used an average of 75 g of perirenal adipose tissue to isolate prASCs, yielding 6.9 × 108 cells seeded in total, corresponding to approximately 9.2 × 106 primary isolated cells per gram tissue
Summary
Mesenchymal stromal/stem cells (MSCs) are immature multipotent stromal cells, which represent a rare population in the perivascular niche within fully specialized tissues throughout theCells 2019, 8, 1346; doi:10.3390/cells8111346 www.mdpi.com/journal/cellsCells 2019, 8, 1346 whole body [1]. Cultured MSCs release a broad range of growth factors, cytokines, and chemokines [4,5] in the culture supernatant that may improve regeneration in injured cells, organs or tissue. The organ-protective effects of MSCs, their conditioned medium or extracellular vesicles have been investigated in the last decade, demonstrating that either infused stem cells or their derivates facilitated tissue and organ regeneration predominantly by released regeneration-promoting factors. MSCs have immunological properties, including anti-inflammatory, immunoregulatory and immunosuppressive capacities [6,7], and are, active immunomodulators during inflammation and sepsis. This immunomodulatory activity has been attributed to the secretion of soluble factors. The TLRs act as sensors for invading pathogens broadly distributed on immune cells and are involved in the pathogenesis of chronic inflammatory and infectious diseases [10]
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