Li Xiaoxia, Xie Jing, Zhai Yue, Fang Tengjiaozi, Rao Nanquan, Hu Shuang, Yang Liping, Zhao Yuming, Wang Yixiang, Ge Lihong
Department of Pediatric Dentistry, Peking University School and Hospital of Stomatology, National Engineering Laboratory for Digital and Material Technology of Stomatology, and Beijing Key Laboratory of Digital Stomatology, Beijing 100081, China.
Department of Stomatology, Shenzhen Children's Hospital, Shenzhen 518026, China.
Stem Cells Int. 2019 Feb 14;2019:2562981. doi: 10.1155/2019/2562981. eCollection 2019.
Retinal degeneration is characterized by the progressive loss of photoreceptors, and stem cell therapy has become a promising strategy. Many studies have reported that mesenchymal stem cell transplantation can sustain retinal structure and prolong retinal functions based on two mechanisms. One is cell replacement, and the other is the paracrine action of stem cells. Cells from human exfoliated deciduous teeth (SHEDs) show characteristics typical of mesenchymal stem cells. They are derived from the neural crest and are a potential cellular source for neural regeneration in stem cell therapy. In this study, we explored the potential of SHEDs to be induced towards the retinal photoreceptor phenotype and to be sustainable in an animal model of retinal degeneration. A factor-cocktail protocol was used to induce SHEDs towards retinal photoreceptors for 24 days, and the characteristics of the induced cells were identified in terms of morphological changes, biomarker expression and subcellular distribution, and calcium influx. SHEDs were labeled with firefly luciferase for tracking by bioluminescent imaging and then transplanted into the subretinal space of mice. Our results showed that SHEDs successfully transdifferentiated into photoreceptor-like cells, which displayed neuron-like morphology, and expressed specific genes and proteins associated with retinal precursors, photoreceptor precursors, and mature photoreceptors. In addition, calcium influx was significantly greater in the retinal-induced than in noninduced SHEDs. tracking confirmed at least 2 weeks of good survival by bioluminescent imaging and 3 months of sustainability of SHEDs by histological analysis. We conclude that SHEDs have the potential to transdifferentiate into retinal photoreceptor-like cells and maintain good viability after transplantation into mice with a normal immune system. This demonstrates preliminary success in generating photoreceptor-like cells from SHEDs and applying SHEDs in treating retinal degeneration.
视网膜变性的特征是光感受器逐渐丧失，而干细胞疗法已成为一种有前景的策略。许多研究报告称，间充质干细胞移植可基于两种机制维持视网膜结构并延长视网膜功能。一种是细胞替代，另一种是干细胞的旁分泌作用。人脱落乳牙干细胞（SHEDs）表现出间充质干细胞的典型特征。它们源自神经嵴，是干细胞疗法中神经再生的潜在细胞来源。在本研究中，我们探讨了SHEDs在视网膜变性动物模型中诱导分化为视网膜光感受器表型并可持续存在的潜力。采用因子鸡尾酒方案将SHEDs诱导为视网膜光感受器24天，并从形态变化、生物标志物表达、亚细胞分布和钙内流等方面鉴定诱导细胞的特征。用萤火虫荧光素酶标记SHEDs以便通过生物发光成像进行追踪，然后将其移植到小鼠的视网膜下间隙。我们的结果表明，SHEDs成功转分化为光感受器样细胞，这些细胞呈现神经元样形态，并表达与视网膜前体、光感受器前体和成熟光感受器相关的特定基因和蛋白质。此外，视网膜诱导的SHEDs中的钙内流明显大于未诱导的SHEDs。生物发光成像追踪证实至少2周内SHEDs存活良好，组织学分析证实SHEDs可持续3个月。我们得出结论，SHEDs有潜力转分化为视网膜光感受器样细胞，并在移植到具有正常免疫系统的小鼠后保持良好的活力。这证明了从SHEDs生成光感受器样细胞并将SHEDs应用于治疗视网膜变性方面取得了初步成功。
