In vitro Differentiation of Adult Adipose Mesenchymal Stem Cells into Retinal Progenitor Cells

Abstract

Introduction: It has been previously shown that adult mesenchymal stem cells (MSCs) differentiate into neural progenitor cells (NPCs) and that the differentiation process was completed in 24–48 h. In this previous study, MSCs from a bone marrow or fat source were co-incubated with homologous autoaggressive cells (ECs) against nerve tissue, and these NPCs were successfully used in human regenerative therapeutic approaches. The present study was conducted to investigate whether a similar differentiation method could be used to obtain autologous retinal progenitor cells (RPCs). Methods: Human Th1 cells against retinal tissue were obtained by challenging human blood mononuclear cells with an eye lysate of bovine origin; negative selection was performed using a specific immunomagnetic bead cocktail. Fat MSCs were obtained from a human donor through mechanical and enzymatic dissociation of a surgical sample. The ECs and MSCs were co-cultured in a serum-free medium without the addition of cytokines for 0, 24, 48 and 72 h. The plastic adherent cells were morphologically examined using inverted-phase microscopy and characterized by immunofluorescent staining using antibodies against Pax 6, TUBB3, GFAP, Bestrophin 2, RPE 65, OPN1 SW, and rhodopsin antigens. Results: The early signs of MSC differentiation into RPCs were observed at 24 h of co-culture, and the early differentiated retinal linage cells appeared at 72 h (neurons, rods, Müller cells, retinal ganglion cells and retinal pigmented epithelial cells). These changes increased during further culture. Conclusion: The results reported here support the development of a method to obtain a large number of autologous adult RPCs, which could be used to treat different retinopathies.

In vitro extracorporeal shock wave treatment enhances stemness and preserves multipotency of rat and human adipose-derived stem cells

“…Several studies have shown that these cells are capable of differentiating into numerous cell types, and differentiation is not restricted to their initial tissue type [9,38,39]. One of the major downsides of ASCs represents their high variability and loss of differentiation capacity at higher passage numbers after expansion in vitro [15,40,41].…”

Comparative study between amniotic-fluid mesenchymal stem cells and retinal pigmented epithelium (RPE) stem cells ability to differentiate towards RPE cells

“…Another approach for RPE replacement foresees the use of mesenchymal stem cells, opportunely differentiated into RPE cells (Duan et al 2013;Moviglia et al 2012). The amniotic fluid may represent an interesting source, since it contains AF-MSCs characterized by a high proliferative and differentiative potential, also towards neurogenic lineages.…”

Differentiation of Mesenchymal Stem Cells into Retinal Progenitor Cells

“…To sum up, the findings of Moviglia et al [1] fall short of providing evidence for their conclusions, for the following reasons:…”

“…We read the paper by Moviglia et al [1] entitled ‘In vitro differentiation of adult adipose mesenchymal stem cells into retinal progenitor cells', published in Opthalmic Research, with great interest.…”

“…The cells isolated by Moviglia et al [1], as described in that paper, only fulfill one of these criteria. The authors do not mention detection of CD105, CD73 or CD90 as positive MSC markers, in the context of negative CD45, CD34, CD14 or CD11b, CD79a or CD19 and HLA class II.…”