Fayazi Mehri, Salehnia Mojdeh, Ziaei Saeideh
Anatomy Department, Tarbiat Modares University, P.O. Box 14115-111, Tehran, Iran.
In Vitro Cell Dev Biol Anim. 2015 Apr;51(4):408-14. doi: 10.1007/s11626-014-9842-2. Epub 2014 Dec 17.
The aim of this study was to investigate the potential differentiation of CD146(+) endometrial stem cells to several lineages. Endometrial stromal cells were cultured using Dulbecco's modified Eagle's medium/Hams F-12 (DMEM/F-12) and were passaged every 7-10 d when cultures reached 80-100% of confluency. The immunophenotypes of single endometrial cells were analyzed using flow cytometry at fourth passage. Then the CD146(+) cells were sorted using magnetic-activated cell sorting, and they were cultured and analyzed for in vitro differentiation to several lineages. Detection of adipocyte- and osteocyte-like cells were assessed by oil red O and alizarin red staining, respectively. For detection of neural progenitor and oligodendrocyte-like cells, the cells were immunostained by neurofilament 68 and oligo2, respectively. The rates of CD90, CD105, CD146, CD31, CD34, and CD9 of cultured endometrial cells were 94.98 ± 3%, 95.77 ± 2.5%, 27.61 ± 2%, 0.79 ± 0.05%, 1.43 ± 0.1%, and 1.01 ± 0.06%, respectively. CD146(+) cells were isolated to high purity. CD146(+)-differentiated cells to adipogenic cell with typical lipid-rich vacuoles and osteogenic cells were observed and confirmed their mesenchymal origin. They also differentiated into neural progenitor and glial differentiation by retinoic acid, basic fibroblast growth factor, and epidermal growth factor signaling molecules, respectively, and confirmed by neurofilament 68 and oligo2 immunocytochemistry. The efficiency of differentiation to neural progenitor and oligodendrocyte-like cells was 90 ± 3.4% and 79 ± 2.8%, respectively. This study showed that CD146(+) cells from human endometrium after in vitro cultivation can differentiate into adipogenic-, osteogenic-, neural progenitor-, and glial-like cells. They may provide available alternative source of stem cells for future cell-based therapies and tissue engineering applications.
本研究的目的是探究CD146(+)子宫内膜干细胞向多个谱系分化的潜能。使用杜氏改良伊格尔培养基/哈姆F-12（DMEM/F-12）培养子宫内膜基质细胞，当培养物达到80-100%汇合度时，每7-10天传代一次。在第4代时使用流式细胞术分析单个子宫内膜细胞的免疫表型。然后使用磁珠分选法分选CD146(+)细胞，并对其进行培养，分析其向多个谱系的体外分化情况。分别通过油红O染色和茜素红染色评估脂肪细胞样细胞和成骨细胞样细胞的检测情况。对于神经祖细胞和少突胶质细胞样细胞的检测，分别用神经丝68和少突胶质细胞转录因子2对细胞进行免疫染色。培养的子宫内膜细胞中CD90、CD105、CD146、CD31、CD34和CD9的表达率分别为94.98±3%、95.77±2.5%、27.61±2%、0.79±0.05%、1.43±0.1%和1.01±0.06%。CD146(+)细胞被分离到高纯度。观察到CD146(+)分化细胞形成了具有典型富含脂质空泡的脂肪生成细胞和成骨细胞，并证实了它们的间充质来源。它们还分别通过视黄酸、碱性成纤维细胞生长因子和表皮生长因子信号分子分化为神经祖细胞和神经胶质细胞，并通过神经丝68和少突胶质细胞转录因子2免疫细胞化学进行了证实。向神经祖细胞和少突胶质细胞样细胞的分化效率分别为90±3.4%和79±2.8%。本研究表明，体外培养后的人子宫内膜CD146(+)细胞可分化为脂肪生成、成骨、神经祖细胞和神经胶质样细胞。它们可能为未来基于细胞的治疗和组织工程应用提供可用的干细胞替代来源。
