Benjamín León-Mancilla , Moisés Martínez-Castillo , Rocío Guerrero , Juan J Montesino , Erika Hernández-Estévez , Maria Cristina Piña-Barba , Rubén S Argüero , Gabriela Gutierrez-Reyes*
Human mesenchymal stem cells (hMSCs) are considered an ideal strategy for tissue engineering and regenerative medicine. However, their acquisition, administration route, and cell quantity are true challenges. On the other hand, the collagen scaffolds are a viable option, mimicking the extracellular matrix, utilizing collagen as the main polymer in 3D supports. The combination of hMSCs and scaffolds could enable the hMSCs to arrive at the target organ, avoiding the disadvantages of intravenous or intra-arterial therapy. We obtain and characterize MSCs from human amniotic membranes and evaluate their differentiation capacity in 3D collagen matrix scaffolds (CMSs). Their morphology, multipotency genes by RT-PCR and markers by flow cytometry were evaluated in in vitro cell cultures. The differentiation capacity of AM-hMSCs, seeded with and without CMSs, was evaluated in media specific for chondrogenic, osteogenic, and adipogenic lineages. AM-hMSCs were studied up to passage 5 and fibroblastoid morphology was observed in AM-hMSCs and BM-hMSCs. Sox2 gene expression was similar in all passages, whereas oct4 was upregulated at P2 and P5. Nanog was upregulated at P1 and P3 versus BM-hMSCs. Membrane markers displayed CD44, CD73, CD90, and CD105 were positive in all passage. AM-MSCs were adhered to CMSs, showing fibroblastoid morphology in the SEM analyses. The AMhMSCs, seeded with and without CMSs, were able to differentiate into chondroblasts, osteoblasts, and adipocytes. The CMSs enable AM-MSC stemness preservation, without affecting their differentiation capacity. That combination can be a novel strategy in the tissue regeneration process.
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