Vossmerbaeumer Urs, Ohnesorge Stefanie, Kuehl Sandra, Haapalahti Minna, Kluter Harald, Jonas Jost B, Thierse Hermann-Josef, Bieback Karen
Department of Ophthalmology, University Eye Hospital, University of Heidelberg, Mannheim, Germany.
Cytotherapy. 2009;11(2):177-88. doi: 10.1080/14653240802714819.
The non-exudative form of age-related macular degeneration (ARMD) is characterized by a progressive decay of retinal pigment epithelium cells at the posterior pole of the eye. As mesenchymal stromal cells (MSC) have been shown to differentiate into various cell types from the mesodermal and ectodermal lineages, we investigated whether we can induce a phenotype displaying retinal pigment epithelium (RPE) characteristics.
The differentiation of human lipo-aspirate-derived MSC toward the RPE lineage was triggered by exposure to conditioned medium from either human or porcine RPE cells. In a second approach we tested whether adding vasoactive intestinal peptide (VIP) is capable of further modifying differentiation processes. Resulting cell populations were assessed for expression of RPE-specific markers by immunofluorescence, quantitative real time (RT)-polymerase chain reaction (PCR) and Western blotting. The potential for pigment synthesis was assessed by the response to melanocyte-stimulating hormone (MSH).
Following culture of undifferentiated MSC with RPE-conditioned medium and/or VIP, expression of typical RPE markers bestrophin, cytokeratins 8 and 18 and RPE 65 was induced. MSH induced the formation of pigmented granula in differentiated MSC.
MSC are shown to express RPE markers upon induction with either RPE-conditioned medium and/or VIP. The gain of basic functional features of RPE cells was indicated by melanin synthesis. This alludes to a differentiation potential of MSC into the neuroectodermal lineage, yielding cells with phenotypic characteristics of RPE cells.
年龄相关性黄斑变性（ARMD）的非渗出型特征为眼后极部视网膜色素上皮细胞的进行性衰退。由于间充质基质细胞（MSC）已被证明可分化为中胚层和外胚层谱系的多种细胞类型，我们研究了是否能够诱导出具有视网膜色素上皮（RPE）特征的表型。
通过暴露于来自人或猪RPE细胞的条件培养基来触发人脂肪抽吸物来源的MSC向RPE谱系的分化。在第二种方法中，我们测试了添加血管活性肠肽（VIP）是否能够进一步改变分化过程。通过免疫荧光、定量实时（RT）-聚合酶链反应（PCR）和蛋白质印迹法评估所得细胞群体中RPE特异性标志物的表达。通过对促黑素细胞激素（MSH）的反应评估色素合成的潜力。
将未分化的MSC与RPE条件培养基和/或VIP一起培养后，诱导了典型RPE标志物Bestrophin、细胞角蛋白8和18以及RPE 65的表达。MSH诱导分化的MSC中形成色素颗粒。
结果表明，用RPE条件培养基和/或VIP诱导后，MSC表达RPE标志物。黑色素合成表明RPE细胞获得了基本功能特征。这暗示了MSC向神经外胚层谱系的分化潜力，产生具有RPE细胞表型特征的细胞。
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