Mouse retinal progenitor cell (RPC) cocultivation with retinal pigment epithelial cell culture affects features of RPC differentiation

Abstract

We provide evidence that coculturing of retinal progenitor cells (RPC) with retinal pigment epithelial cells significantly biases the standard in vitro RPC differentiation patterns. In particular, in cocultivation experiments RPCs lost the ability to differentiate spontaneously and displayed approximately 2.1-2.4-fold increase in immunoreactivity to the neural stem cell marker nestin and approximately 1.6-1.7-fold increase in rod photoreceptor cell rhodopsin marker immunoreactivity. The data suggest the influence of the intercellular interaction networks on RPC differentiation.

Regulation of Prenatal Human Retinal Neurosphere Growth and Cell Fate Potential by Retinal Pigment Epithelium and Mash1

“…Taken together, our findings suggest that the fate potential of long‐term retinal neurosphere cultures is governed in large part by intrinsic limitations rather than extrinsic influences. Consistent with this hypothesis, culture modifications that enhanced neurogenesis in rodent RPCs failed to alter cell fate potential in late‐passage human retinal neurospheres [66, 68, 69]. Therefore, we misexpressed the proneural bHLH transcription factor Mash1 in an attempt to overcome Hes signaling and push the hRPCs toward a neuronal fate.…”

“…In an attempt to re‐establish the potential of long‐term retinal neurosphere cultures to produce neurons, we first tried varying the differentiation conditions. However, changing the cell plating density [66, 67] or exposing differentiating cells to selected factors (e.g., FGF2 or NT4) [50, 68, 69–70] did not alter the fate potential (data not shown). Since manipulation of the cell environment was unsuccessful, we next sought to modify the intrinsic gene expression profile by misexpressing Mash1, a proneural bHLH transcription factor known to play an important role in the differentiation of multiple neuronal cell types within the brain and retina, including bipolar cells and photoreceptors [11, 71, 72, 73, 74, 75–76].…”

Treatment viability of stem cells in ophthalmology

“…Humans, unlike lower vertebrates, exhibit little RSC activity in adulthood resulting in limited capacity for retinal regeneration [10]. Multipotent progenitor (nestin positive) cells have now been isolated from the brain and retina of postnatal chick and the adult mouse expanded in culture, and transplanted to the CNS [34,35]. Progenitor cells derived from the CNS readily engrafted in the diseased retina of mature recipients, where they developed morphologies appropriate to the local microenvironment and expressed mature markers such as photoreceptor protein rhodopsin [36].…”

Caracterización de los progenitores neurales de retina adulta del pez cebra

“…Además de la retirada de los factores de crecimiento, se han realizado diversos ensayos añadiendo distintas moléculas para dirigir la diferenciación celular. Entre ellas, además del FGF ya mencionado, se ha testado el suero fetal bobino Liu et al, 2005;MacNeil et al, 2007;Kokkinopoulos et al, 2008), eĺ acido retinóico (Stenkamp et al, 1993;Kelley et al, 1999;Lawrence et al, 2007;Martínez-Monedero et al, 2008;Osakada et al, 2008;Safari et al, 2009;Wu et al, 2012), la proteína Shh (Martínez-Monedero et al, 2008;Zeng et al, 2007;Osakada et al, 2008;Osterberg y Roussa, 2009;Wu et al, 2012) o un medio condicionado derivado del epitelio pigmentario (Gaur et al, 1992;Kholodenko et al, 2006;Sheedlo et al, 2007;Gamm et al, 2008).…”