Purpose
Human amniotic membrane-derived mesenchymal cells (hAMCs) has been shown the potential of three germ differentiations, and may be expected to show immunologic tolerance. In this study, we try to optimize differentiation protocols for inducing and maintaining directed differentiation and gain functional characteristics of induced retinal neuron from hAMCs, with regard to their morphology, survival, migration and differentiation potential in vivo.
Methods
hAMCs were ex vivo expanded from human amnionic membrane (HAM), and CD73, CD90, CD105, CD44, CD166, and CD45 expression was assessed by flow cytometry. Their Mesodermal differentiation was measured by osteogenesis and adipogenesis. Normal PCR and real-time PCR analysis were used to assess OCT-4, NANOG, REX-1, SOX-2, SSEA-4 and vimentin expression in hAMCs. Induced hAMCs were grafted to ventriculus lateralis of rats and observed. Nestin, Map2, Ngn2, and Visinin were detected in transplanted rats by immunohistochemistry and immunocytochemistry.
Results
hAMCs express typical mesenchymal marker (CD73, CD90, CD105), and displayed strong adipogenic and osteogenic differentiation potential. And we induced hAMCs conversion of retinal neural like cells in conditional medium with 4-stage protocol. The induced cells can be labeled retinal neuron progenitor and mature cell markers (nestin, MAP2, ngn2 and visinin). These cells were then transplanted into the Lateral ventricle of rats, which survived and migrated in the ventricle of rats, and can be labeled with ngn2 and visinin.
Conclusions
hAMCs can be induced to retinal neuron-like cells; this may be a promising cell resource in clinical treatment for retinal disease.
Keywords: 721 stem cells •
741 transplantation •
688 retina