Article
Gene interaction profiles of differentiated and undifferentiated adipose derived mesenchymal stem cells dynamically seeded onto a processed nerve allograft
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Published: | February 6, 2020 |
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Objectives/Interrogation: The gene expression profile of adipose derived mesenchymal stem cells (MSCs) seeded onto a substrate varies depending on the cells interaction with the remaining extracellular matrix. The effect of undifferentiated cells or cells differentiated into Schwann-like cells seeded onto a processed nerve allograft is unknown. The purpose of this study was to determine the gene expression profiles of differentiated and undifferentiated MSCs before and after seeding onto a processed nerve allograft in order to determine the mechanism of action of both stem cell types when used in peripheral nerve repair.
Methods: 65 sciatic nerve segments from Sprague-Dawley rats were harvested and decellularized. MSCs were harvested from Lewis rats and cultured. Half of the MSCs were differentiated into Schwann-like cells. Undifferentiated MSCs and differentiated MSCs were dynamically seeded on the surface of the decellularized allografts. The interaction between cells and the nerve graft was evaluated by qPCR-analysis of neurotrophic (GDNF, PTN, GAP43, PMP22), angiogenic (CD31, VEGF1), extracelullar matrix (ECM) (COL1A1, COL3A1, FBLN1, LAMB2) and regulatory cell cycle genes (CAPS3, CCBN2).
Results and Conclusions: Seeding of undifferentiated MSCs led to enhanced expressions of neurotrophic (NGF, GDNF, PMP22), ECM (FBLN1, LAMB2) and regulatory cell cycle genes (CCNB2), mostly after 7 days, while the interaction with differentiated MSCs led to enhanced expressions of neurotrophic (NGF, GDNF, GAP43), angiogenic (VEGF1), ECM (FBLN1) and regulatory cell cycle genes (CASP3, CCNB2) within the first week.
Undifferentiated and differentiated MSCs have a different baseline gene expression, which changes after seeding onto a processed nerve allograft. Differentiated MSCs were observed to express high regenerative gene levels in the first 72 hours after seeding, while undifferentiated MSCs expressed high regenerative gene levels a week after seeding. These findings imply that both cell-types effect nerve regeneration in different stages and that a combination of undifferentiated AMSCs and differentiated AMSCs might lead to superior nerve regeneration.