通过一种简单的方法，在含有小分子诱导剂的培养基中高效地将人脂肪来源干细胞分化为视网膜色素上皮样细胞。
High efficient differentiation of human adipose-derived stem cells into retinal pigment epithelium-like cells in medium containing small molecules inducers with a simple method.
作者信息
Aboutaleb Kadkhodaeian Hamid, Salati Amir, Lashay Alireza
机构信息
Nervous System Stem Cells Research Center, Semnan University of Medical Sciences, Semnan, Iran; Department of Anatomical Sciences, Faculty of Medicine, Semnan University of Medical Sciences, Semnan, Iran.
Nervous System Stem Cells Research Center, Semnan University of Medical Sciences, Semnan, Iran; Department of Tissue Engineering and Applied Cell Sciences, Faculty of Medicine, Semnan University of Medical Sciences, Semnan, Iran.
出版信息
Tissue Cell. 2019 Feb;56:52-59. doi: 10.1016/j.tice.2018.12.003. Epub 2018 Dec 12.
BACKGROUND
The induction of retinal pigmented epithelium cells (RPE) is one of the most important objectives in research focused on treating retinal degenerative diseases. The present study aims to differentiate human adipose stem cells (hADSCs) into RPE cells for replacement therapies in cases of retinal degenerative diseases.
METHODS
Lipoaspirate-derived human adipose stem cells (LA-hADSCs) were obtained from abdominal samples and examined by immunocytochemistry for the expression of mesenchymal adipose stem cell markers. RPE cells were also obtained from human samples and cultured to be used as control after being examined for the expression of their designated markers. hADSCs differentiated into RPE cells after 80 days using chemical inducers in one steps. The differentiated cells were then compared to control cells in marker expression. The differentiated cells were also examined under a scanning electron microscope for the presence of apical microvilli and cell connection.
RESULTS
Cultured hADSCs at the fourth passage was shown to express the surface markers CD90 (98 ± 2%), CD11b (96 ± 3%), and CD105 (95 ± 4%). The RPE cells obtained from human samples expressed the marker RPE65 quite well. 80 days after differentiation, the previously hADSCs expressed both RPE65 (100%) and CRALBP (96 ± 1%) and were thus significantly similar to the RPE cells obtained from human samples. Morphologically, differentiated cells appeared to have epithelial and cytoplasmic pigment granules. Observations using a scanning electron microscope recorded clear connections among the differentiated RPE cells and revealed apical microvilli.
CONCLUSION
Human adipose stem cells can differentiate into retinal pigmented epithelium cells, which can be used in cell replacement therapy for degenerative diseases including age-related macular degeneration (AMD) as well as retinitis pigmentosa (RP).
背景
诱导视网膜色素上皮细胞（RPE）是专注于治疗视网膜退行性疾病研究的最重要目标之一。本研究旨在将人脂肪干细胞（hADSCs）分化为RPE细胞，用于视网膜退行性疾病的替代疗法。
方法
从腹部样本中获取抽脂来源的人脂肪干细胞（LA-hADSCs），通过免疫细胞化学检测间充质脂肪干细胞标志物的表达。还从人样本中获取RPE细胞，在检测其指定标志物的表达后进行培养用作对照。hADSCs在80天内使用化学诱导剂一步分化为RPE细胞。然后将分化后的细胞与对照细胞进行标志物表达比较。还在扫描电子显微镜下检查分化后的细胞是否存在顶端微绒毛和细胞连接。
结果
培养至第4代的hADSCs显示表达表面标志物CD90（98±2%）、CD11b（96±3%）和CD105（95±4%）。从人样本中获得的RPE细胞很好地表达了标志物RPE65。分化80天后，先前的hADSCs表达了RPE65（100%）和CRALBP（96±1%），因此与从人样本中获得的RPE细胞显著相似。在形态上，分化后的细胞似乎具有上皮和细胞质色素颗粒。使用扫描电子显微镜观察记录到分化后的RPE细胞之间有清晰的连接，并显示有顶端微绒毛。
结论
人脂肪干细胞可分化为视网膜色素上皮细胞，可用于包括年龄相关性黄斑变性（AMD）以及视网膜色素变性（RP）在内的退行性疾病的细胞替代治疗。
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