In vitro differentiation of retinal ganglion-like cells from embryonic stem cell derived neural progenitors

Molecular Control of Retinal Ganglion Cell Specification and Differentiation

“…In the post-hatch chicken retina, FGF signaling is required to induce Atoh7 expression and subsequent production of Pou4f1-and Isl1-immunoreactive RGCs from stem cells in the ciliary marginal zone (Fischer et al, 2002). Similarly, mouse embryonic stem (ES) cells can be induced to generate RGC-like cells that express Atoh7, Pou4f2 and Isl1 only when exposed to FGF2, and this RGC-inducing activity by FGF2 is further potentiated by Shh (Jagatha et al, 2009). In rat retinal progenitors, transient expression of the intrinsic factor Atoh7 led to Pou4f2 upregulation and RGC induction (Yao et al, 2007).…”

“…Conceivably, application of a combination of stimulatory intrinsic and extrinsic factors while suppressing negative ones may lead to even greater induction of RGCs from stem cells. Transplanting in vitro induced retinal progenitors and RGCs intravitreally into postnatal rodent eyes resulted in only few cells that integrated into the RGC layer of the host retina (Chen et al, 2010;Jagatha et al, 2009), suggesting that postnatal retinas present a formidable barrier to cell migration and integration.…”

Modeling autosomal dominant optic atrophy using induced pluripotent stem cells and identifying potential therapeutic targets

“…iPSC technology circumvents these issues since patient-specific iPSCs can be generated, and these iPSCs have the ability to differentiate into various cell types [23,36,37]. It has also been reported recently that retinal ganglionlike cells can be generated from iPSCs [28,[38][39][40]. Several protocols have been developed and published for the retina-specific differentiation of stem cells.…”

Neuronal Differentiation of Embryonic Stem Cell Derived Neuronal Progenitors Can Be Regulated by Stretchable Conducting Polymers

“…RA induced EBs were partially differentiated on poly-D-lysine (150 mg/mL) and laminin (1 mg/mL) coated substrate for 2 days in N2 differentiation medium (DMEM/ F12) supplemented with 1% N2 supplement, FGF2 (10 ng/ mL), or epidermal growth factor (EGF) (10 ng/mL), heparin (2 mg/mL), and 0.5% FBS. 39 The partially differentiated EBs were trypsinized mildly and plated onto uncoated six-well plates (*1.5 · 10 6 cells/well) in proliferation medium consisting of DMEM/F12 supplemented with 1% N2 supplement, heparin (2 mg/mL), and FGF2 (20 ng/mL) and allowed to proliferate further for 4 days.…”

“…39 Briefly, 4% paraformaldehydefixed cells were blocked in 5% NGS (Sigma-Aldrich) and permeabilized with 0.2%-0.4% Triton-X 100 followed by an overnight incubation with primary antibodies at 4°C (b-III tubulin 1:200 Chemicon, GFAP 1:400 Sigma). Cells were examined for fluorescence after incubation with appropriate secondary antibody conjugated to FITC (1:400) or cy3 (1:400).…”