Monnin Julie, Morand-Villeneuve Nadège, Michel Germaine, Hicks David, Versaux-Botteri Claudine
Laboratoire de Neurosciences, Université de Franche-Comté EA 481, IFR133, Place du Maréchal Leclerc, 25030 Besançon, Cedex, France.
Neurosci Lett. 2007 Jun 21;421(1):22-6. doi: 10.1016/j.neulet.2007.04.073. Epub 2007 May 16.
In lower vertebrates, like fishes and amphibians, retina is able to self-regenerate whereas Mammalian retina has lost this property. Nevertheless, recently, it has turned out that retinal glial Müller cells were playing a role in neuronal regeneration of the adult rodent retina, in case of acute damages, by dedifferentiating and redifferentiating in glial and neural cells. The purpose of this study was to analyse the ability of mammalian Müller cells for forming neurospheres. First of all, rats Müller cells were isolated in a primary culture. Second, these cells were resuspended in two different culture media: the cells cultures in the Neurobasal-A medium kept a typical Müller cells morphology even after 15 days of EGF treatment, and the cells plated in the DMEM-F12 medium formed neurospheres from the third day in culture. The neurosphere cells expressed nestin, cellular retinaldehyde binding protein (CRALBP) and glial fibrillary acidic protein (GFAP). These results showed our capacity to isolate and propagate Müller cells-derived progenitor cells. Moreover, it allows us to control the number of progenitor cells and, in the future, to study their differentiation capacity.
在鱼类和两栖类等低等脊椎动物中，视网膜能够自我再生，而哺乳动物的视网膜已丧失这一特性。然而，最近发现，在成年啮齿动物视网膜急性损伤的情况下，视网膜神经胶质穆勒细胞通过去分化并重新分化为神经胶质细胞和神经细胞，在神经元再生中发挥作用。本研究的目的是分析哺乳动物穆勒细胞形成神经球的能力。首先，在原代培养中分离大鼠的穆勒细胞。其次，将这些细胞重悬于两种不同的培养基中：在添加表皮生长因子（EGF）处理15天后，在Neurobasal - A培养基中培养的细胞仍保持典型的穆勒细胞形态，而接种于DMEM - F12培养基中的细胞从培养第三天开始形成神经球。神经球细胞表达巢蛋白、细胞视黄醛结合蛋白（CRALBP）和胶质纤维酸性蛋白（GFAP）。这些结果表明我们有能力分离和扩增源自穆勒细胞的祖细胞。此外，这使我们能够控制祖细胞的数量，并在未来研究它们的分化能力。
