Human Bone Marrow Stromal Cells can Differentiate to a Retinal Pigment Epithelial Phenotype when Co-Cultured with Pig Retinal Pigment Epithelium using a Transwell System

Abstract

Background: There is an increasing interest in generating retinal pigment epithelial (RPE) cells from stem cells for therapy against degenerative eye diseases. Human bone marrow stromal cells (hBMSCs) can be induced to express retinal neuron-specific markers when co-cultured with retinal neurons, however, whether hBMSCs can differentiate into RPE-like cells in a co-culture system has not been clarified. Methods: The induction of hBMSCs into RPE-like cells was performed by combining hBMSCs and pig RPE cells in a transwell system. The biomarkers of hBMSCs-derived RPE cells were determined by quantitative RT-PCR and immunofluorescence. The function of induced cells was assayed by ELISA for secretion of neurotrophic factors. Results: Intracellular pigment granules and many RPE markers existed in hBMSCs-derived RPE cells after co-culturing with pig RPE cells for 14 days. Typical RPE functions, such as phagocytosis of photoreceptor outer segments and secretion of the trophic factors, brain-derived neurotrophic factor (BDNF) and glia-derived neurotrophic factor (GDNF), were observed in these induced cells. Conclusion: hBMSCs can be induced toward functional RPE cells simply by transwell-based co-culture with RPE cells.

Stem cell based therapies for age-related macular degeneration: The promises and the challenges

“…A 32-gauge blunt cannula, Hamilton syringe (Hamilton, Reno, NV), is used to inject the cell suspension Lu et al, 2009;Diniz et al, 2013;Hu et al, 2012). In terms of the amount of cells injected in each eye, previous studies have described different quantities that vary from 5 Â 10 3 to 1 Â 10 5 cells/eye in RCS rats Lu et al, 2009;Duan et al, 2013). When the choroid is dissected 1e2 weeks after the injection, a small, localized hemorrhage is usually seen near the site of injection.…”

c-Kit+ cells isolated from human fetal retinas represent a new population of retinal progenitor cells

“…Immunocytochemistry was performed using our previously described methods (Duan et al, 2013). Briefly, the cells or slides were fixed and permeabilized, then blocked in 10% serum for 30 min.…”

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.…”