Pluripotency and immunomodulatory signatures of canine induced pluripotent stem cell-derived mesenchymal stromal cells are similar to harvested mesenchymal stromal cells

Abstract

With a view towards harnessing the therapeutic potential of canine mesenchymal stromal cells (cMSCs) as modulators of inflammation and the immune response, and to avoid the issues of the variable quality and quantity of harvested cMSCs, we examined the immunomodulatory properties of cMSCs derived from canine induced pluripotent stem cells (ciMSCs), and compared them to cMSCs harvested from adipose tissue (cAT-MSC) and bone marrow (cBM-MSC). A combination of deep sequencing and quantitative RT-PCR of the ciMSC transcriptome confirmed that ciMSCs express more genes in common with cBM-MSCs and cAT-MSCs than with the ciPSCs from which they were derived. Both ciMSCs and harvested cMSCs express a range of pluripotency factors in common with the ciPSCs including NANOG, POU5F1 (OCT-4), SOX-2, KLF-4, LIN-28A, MYC, LIF, LIFR, and TERT. However, ESRRB and PRDM-14, both factors associated with naïve, rather than primed, pluripotency were expressed only in the ciPSCs. CXCR-4, which is essential for the homing of MSCs to sites of inflammation, is also detectable in ciMSCs, cAT- and cBM-MSCs, but not ciPSCs. ciMSCs constitutively express the immunomodulatory factors iNOS, GAL-9, TGF-β1, PTGER-2α and VEGF, and the pro-inflammatory mediators COX-2, IL-1β and IL-8. When stimulated with the canine pro-inflammatory cytokines tumor necrosis factor-α (cTNF-α), interferon-γ (cIFN-γ), or a combination of both, ciMSCs upregulated their expression of IDO, iNOS, GAL-9, HGF, TGF-β1, PTGER-2α, VEGF, COX-2, IL-1β and IL-8. When co-cultured with mitogen-stimulated lymphocytes, ciMSCs downregulated their expression of iNOS, HGF, TGF-β1 and PTGER-2α, while increasing their expression of COX-2, IDO and IL-1β. Taken together, these findings suggest that ciMSCs possess similar immunomodulatory capabilities as harvested cMSCs and support further investigation into their potential use for the management of canine immune-mediated and inflammatory disorders.

Comparative characteristic study from bone marrow-derived mesenchymal stem cells

“…In addition, Rex1 has been considered a pluripotency marker [ 51 ]. It has been reported that pluripotency TFs are expressed in all BM-MSCs derived from human [ 51 52 53 ], canine [ 46 54 ], rat [ 55 ], and murine [ 56 ] sources. Moreover, Ki67 is a proliferation marker, based on its reported expression in BM-MSCs derived from rat [ 55 ] and canine [ 46 ] but not human [ 57 ] sources.…”

Comparative analysis of mesenchymal stem/stromal cells derived from human induced pluripotent stem cells and the cognate umbilical cord mesenchymal stem/stromal cells

“…Current, the immunomodulatory effect of iMSCs which play crucial role in regenerative medicine is still controversial theme [ 26 ]. For example, previous study has showed the immunomodulatory signatures of canine iMSCs are similar to the adipose tissue and bone marrow derived MSCs [ 47 ]. However, some other studies had showed iMSCs has better immunomodulatory effects than conventional MSCs, for example, Schnabel et al, has reported iMSCs had similar immunogenic properties but more potent immunomodulatory effects than tissue source MSCs [ 48 ].…”

Induced pluripotent stem cells in companion animals: how can we move the field forward?

“…Canine iMSCs have been successfully differentiated into osteogenic and chondrogeneic lineage ( 90 , 135 ). Furthermore, from a functional perspective, canine iMSCs show an immunomodulatory capacity similar to primary MSCs derived from adipose tissue and bone marrow, with similar gene expression profiles, effects on the proliferation of T cells, maturation of dendritic cells and response to priming with pro-inflammatory cytokines ( 89 , 136 ). Furthermore, canine iMSCs IV injected in three healthy dogs did not produce adverse events in the short term, neither tumor formation was observed up to 15 months of monitoring ( 89 ).…”