Systems biology analysis of osteogenic differentiation behavior by canine mesenchymal stem cells derived from bone marrow and dental pulp

Abstract

Utilization of canine mesenchymal stem cells (cMSCs) for regenerating incorrigible bone diseases has been introduced. However, cMSCs harvested from different sources showed distinct osteogenicity. To clarify this, comparative proteomics-based systems biology analysis was used to analyze osteogenic differentiation behavior by cMSCs harvested from bone marrow and dental pulp. The results illustrated that canine dental pulp stem cells (cDPSCs) contained superior osteogenicity comparing with canine bone marrow-derived MSCs (cBM-MSCs) regarding alkaline phosphatase activity, matrix mineralization, and osteogenic marker expression. Global analyses by proteomics platform showed distinct protein clustering and expression pattern upon an in vitro osteogenic induction between them. Database annotation using Reactome and DAVID revealed contrast and unique expression profile of osteogenesis-related proteins, particularly on signaling pathways, cellular components and processes, and cellular metabolisms. Functional assay and hierarchical clustering for tracking protein dynamic change confirmed that cBM-MSCs required the presences of Wnt, transforming growth factor (TGF)-beta, and bone-morphogenetic protein (BMP) signaling, while cDPSCs mainly relied on BMP signaling presentation during osteogenic differentiation in vitro. Therefore, these findings illustrated the comprehensive data regarding an in vitro osteogenic differentiation behavior by cBM-MSCs and cDPSCs which is crucial for further mechanism study and the establishment of cMSC-based bone tissue engineering (BTE) for veterinary practice.

Intrathecal Transplantation of Autologous and Allogeneic Bone Marrow-Derived Mesenchymal Stem Cells in Dogs

“…Our flow cytometry assay was able to characterize canine BM-MSCs as CD90 + , CD44 + , CD34 − , CD45 − , e MHC II − . Herein, canine BM-MSCs exhibited a comparable in vitro profile as described in previous studies 39 , 43 – 46 .…”

Comparative characteristic study from bone marrow-derived mesenchymal stem cells

“…Morphological features are important in defining MSCs as they present as fibroblast-like cells. It has been reported that this feature is possessed by BM-MSCs derived from human [ 45 ], canine [ 46 47 ], feline [ 19 ], rat [ 48 ], and murine [ 49 ] sources. To the best of our knowledge, murine MSCs are smaller than human, canine, and feline 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.…”

Tailored generation of insulin producing cells from canine mesenchymal stem cells derived from bone marrow and adipose tissue

“…Their characteristics were similar to those described in previous reports, including fibroblast-like structures, mRNA expressions related to stemness and proliferation, and MSC-related surface marker expression, along with the multilineage differentiation potential toward osteogenic, chondrogenic, and adipogenic lineages 16 – 18 , 31 – 34 . In this study, we employed the flow cytochemistry analysis on the expression of representative MSC-related and hematopoietic surface markers (Cd73, Cd90, and Cd45) according to our recent publication 35 . The expression of Cd73 in both MSCs was relatively low, as mentioned in a previous report 36 .…”

“…Recent publication from our team regarding bone tissue engineering also showed the distinct osteogenic differentiation potential by cBM-MSCs and canine dental pulp stem cells (cDPSCs) in vitro. To dissect the underlying mechanisms, proteomics-based systems biology analysis was applied, and it suggested that both cells required different signaling pathways and underlying mechanisms for regulating their osteogenic paths 35 .…”