Extracellular Nucleotide Hydrolysis in Dermal and Limbal Mesenchymal Stem Cells: A Source of Adenosine Production

Abstract

Human Limbal (L-MSCs) and Dermal Mesenchymal Stem Cell (D-MSCs) possess many properties that increase their therapeutic potential in ophthalmology and dermatology. It is known that purinergic signaling plays a role in many aspects of mesenchymal stem cells physiology. They release and respond to purinergic ligands, altering proliferation, migration, differentiation, and apoptosis. Therefore, more information on these processes would be crucial for establishing future clinical applications using their differentiation potential, but without undesirable side effects. This study evaluated and compared the expression of ecto-nucleotidases, the enzymatic activity of degradation of extracellular nucleotides and the metabolism of extracellular ATP in D-MSCs and L-MSCs, isolated from discard tissues of human skin and sclerocorneal rims. The D-MSCs and L-MSCs showed a differentiation potential into osteogenic, adipogenic, and chondrogenic lineages and the expression of markers CD105 , CD44 , CD14 , CD34 , CD45 , as expected. Both cells hydrolyzed low levels of extracellular ATP and high levels of AMP, leading to adenosine accumulation that can regulate inflammation and tissue repair. These cells expressed mRNA for ENTPD1, 2, 3, 5 and 6, and CD73 that corresponded to the observed enzymatic activities. Thus, considering the degradation of ATP and adenosine production, limbal MSCs are very similar to dermal MSCs, indicating that from the aspect of extracellular nucleotide metabolism L-MSCs are very similar to the characterized D-MSCs. J. Cell. Biochem. 118: 2430-2442, 2017. © 2017 Wiley Periodicals, Inc.

How to influence the mesenchymal stem cells fate? Emerging role of ectoenzymes metabolizing nucleotides

“…The above-described sets of experiments allowed the formulation of a general conclusion that the high 5′-NT activity is characteristic of MSCs presenting a high proliferative potential and susceptibility to the positive, stimulatory effect of Ado. The ecto-NTPDases activity on the surface of these cells is negligible (Iser et al, 2014;Kerkelä et al, 2016;Naasani et al, 2017;Roszek, Bomastek et al, 2013). MSCs from different murine tissues and also showed that these discrepancies are related to differential ecto-NTPDase (CD39) and ecto-5′-NT (CD73) expression profile of these cells.…”

“…It is worth pointing out that, on their surface, MSCs express different ectonucleotidases that metabolize extracellular ATP and other nucleotides, hence modulating the purinergic signaling dynamics (Naasani et al, ; Noronha‐Matos & Correia‐de‐Sá, ; Roszek, Błaszczak et al, ; Roszek, Bomastek et al, ; Roszek et al, ; Scarfì, ).…”

“…The above‐described sets of experiments allowed the formulation of a general conclusion that the high 5′‐NT activity is characteristic of MSCs presenting a high proliferative potential and susceptibility to the positive, stimulatory effect of Ado. The ecto‐NTPDases activity on the surface of these cells is negligible (Iser et al, ; Kerkelä et al, ; Naasani et al, ; Roszek, Bomastek et al, ).…”

Human mesenchymal stromal cells inhibit platelet activation and aggregation involving CD73-converted adenosine

“…In contrast, MSCs had low CD39 and low A2AR expression but high CD73 expression. The observed differences between BM-MSCs, LA-MSCs and CB-MSCs may relate to differences in nucleotide hydrolysis activity [ 32 ]. In contrast to our data, other authors detected CD39 expression in MSCs.…”

“…Ectonucleotidase activity was measured in the cells as described previously [ 32 ]. Briefly, cells were seeded at 10,000 cells/cm 2 in 24-well plates and then incubated for 24 h. ATP, ADP and AMP (1 mM; Santa Cruz) were then added in phosphate-free buffer and incubated for either 1 h (ATP and ADP) or 30 min (AMP).…”