HUMAN DENTAL PULP STEM CELLS RESPOND TO CUES FROM THE RAT RETINA AND DIFFERENTIATE TO EXPRESS THE RETINAL NEURONAL MARKER RHODOPSIN
HUMAN DENTAL PULP STEM CELLS RESPOND TO CUES FROM THE RAT RETINA AND DIFFERENTIATE TO EXPRESS THE RETINAL NEURONAL MARKER RHODOPSIN
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DOI：
10.1016/j.neuroscience.2014.09.023
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发表时间：
2014-11-07
期刊：
影响因子：
3.3
通讯作者：
Lamas, M.
中科院分区：
文献类型：
--
作者：
Bray, A. F.;Cevallos, R. R.;Lamas, M.
人类成人牙髓干细胞（DPSC）是自我更新的干细胞，在发育过程中起源于神经嵴，并在成年期保留在牙髓微环境中。由于它们的多谱系分化潜力和相对容易获得，它们代表了神经退行性疾病中基于自体干细胞的疗法的令人兴奋的替代方案。在动物模型中，移植到大脑中的 DPSC 会分化为功能性神经元或星形胶质细胞，以响应似乎影响存活细胞命运的局部环境线索。在这里，我们测试了这样的假设：DPSC 可能能够对视网膜中存在的因素做出反应，从而使这些细胞具有再生潜力。我们评估了 DPSC 对来自对照和化学损伤大鼠视网膜的器官型外植体的条件培养基的反应。为了评估细胞分化，我们通过免疫荧光和逆转录聚合酶链反应（RT-PCR）分析了胶质纤维酸性蛋白（GFAP）、早期神经元和视网膜标记物（聚唾液酸-神经细胞粘附分子（PSA-NCAM）；Pax6；Ascl1；NeuroD1）和晚期感光标记物视紫红质的表达。将 DPSC 培养物暴露于对照视网膜的条件培养基中，导致表达 GFAP 的 DPSC 数量减少 39%； Pax6、Ascl1、PSA-NCAM 或 NeuroD1 的表达检测不到或没有显着变化。在对照中或将培养物暴露于视网膜条件培养基后，未检测到视紫红质的表达。相比之下，暴露于受损视网膜条件培养基的 DPSC 中，44% 对该蛋白呈免疫阳性。当使用富含 Muller 的原代培养物的条件培养基时，无法重现这种反应。最后，进行定量RT-PCR来比较与视网膜器官型外植体共培养的DPSC中胶质细胞源性神经营养因子（GDNF）、神经生长因子（NGF）、睫状神经营养因子（CNTF）和脑源性神经营养因子（BDNF）的相对表达，其中BDNF mRNA表达在视网膜暴露的培养物中显着上调。我们的数据表明 DPSC 培养物对来自大鼠视网膜的信号做出反应并分化以表达视网膜神经元标记。 (C) 2014 年国际广播组织。由爱思唯尔有限公司出版。保留所有权利。
Human adult dental pulp stem cells (DPSCs) are self-renewing stem cells that originate from the neural crest during development and remain within the dental pulp niche through adulthood. Due to their multi-lineage differentiation potential and their relative ease of access they represent an exciting alternative for autologous stem cell-based therapies in neurodegenerative diseases. In animal models, DPSCs transplanted into the brain differentiate into functional neurons or astrocytes in response to local environmental cues that appear to influence the fate of the surviving cells. Here we tested the hypothesis that DPSCs might be able to respond to factors present in the retina enabling the regenerative potential of these cells. We evaluated the response of DPSCs to conditioned media from organotypic explants from control and chemically damaged rat retinas. To evaluate cell differentiation, we analyzed the expression of glial fibrillary acidic protein (GFAP), early neuronal and retinal markers (polysialic acid-neural cell adhesion molecule (PSA-NCAM); Pax6; Ascl1; NeuroD1) and the late photoreceptor marker rhodopsin, by immunofluorescence and reverse transcription polymerase chain reaction (RT-PCR). Exposure of DPSC cultures to conditioned media from control retinas induced a 39% reduction on the number of DPSCs that expressed GFAP; the expression of Pax6, Ascl1, PSA-NCAM or NeuroD1 was undetectable or did not change significantly. Expression of rhodopsin was not detectable in control or after exposure of the cultures with retinal conditioned media. By contrast, 44% of DPSCs exposed to conditioned media from damaged retinas were immunopositive to this protein. This response could not be reproduced when conditioned media from Muller-enriched primary cultures was used. Finally, quantitative RT-PCR was performed to compare the relative expression of glial cell-derived neurotrophic factor (GDNF), nerve growth factor (NGF), ciliary neurotrophic factor (CNTF) and brain-derived neurotrophic factor (BDNF) in DPSC co-cultured with retinal organotypic explants, where BDNF mRNA expression was significantly upregulated in retinal-exposed cultures. Our data demonstrate that DPSC cultures respond to cues from the rat retina and differentiate to express retinal neuronal markers. (C) 2014 IBRO. Published by Elsevier Ltd. All rights reserved.