Establishment of immortalized mesenchymal stem cells derived from the submandibular glands of tdTomato transgenic mice.
Establishment of immortalized mesenchymal stem cells derived from the submandibular glands of tdTomato transgenic mice.
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DOI：
10.3892/etm.2015.2700
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发表时间：
2015-10
影响因子：
2.7
通讯作者：
Miura H
中科院分区：
文献类型：
--
作者：
Furukawa S;Kuwajima Y;Chosa N;Satoh K;Ohtsuka M;Miura H;Kimura M;Inoko H;Ishisaki A;Fujimura A;Miura H
过表达红色荧光蛋白tdTomato的转基因小鼠（tdTomato小鼠）非常适合用于再生医学研究。来自该小鼠模型的培养细胞显示出强烈的红色荧光，使得能够通过移植动物的身体表面进行实时体内成像。间充质干细胞（MSC）在细胞治疗和再生医学中具有显着的潜力;然而，对其体内动力学的调节机制知之甚少。在本研究中，MSC系来源于tdTomato小鼠的下颌下腺成纤维细胞。在整个身体的器官以及唾液腺中观察到来自该细胞系的荧光信号。来源于下颌下腺的原代培养细胞用SV 40大T抗原（GManSV细胞）永生化;这些细胞表现出增加的迁移能力，与从舌下腺分离的细胞相比。GManSV细胞是tdTomato阳性的，并表现出纺锤形成纤维细胞形态;它们还强烈表达小鼠MSC标志物：干细胞抗原-1（Sca-1）、CD 44和CD 90。该细胞系保留了多能干细胞的特性，如其分化为成骨和成脂谱系的能力所证明的。这些结果表明，Sca-1+/CD 44 +/CD 90 +-GManSV细胞可能是有用的动力学研究下颌下腺来源的骨髓间充质干细胞的背景下，在体外共培养与其他类型的唾液腺来源的细胞。这些细胞也可用于体内成像研究，以确定用于治疗唾液腺疾病的新型细胞疗法和再生医学。
Transgenic mice that overexpress the red fluorescent protein tdTomato (tdTomato mice) are well suited for use in regenerative medicine studies. Cultured cells from this murine model exhibit strong red fluorescence, enabling real-time in vivo imaging through the body surface of grafted animals. Mesenchymal stem cells (MSCs) have marked potential for use in cell therapy and regenerative medicine; however, the mechanisms that regulate their dynamics in vivo are poorly understood. In the present study, an MSC line was derived from the submandibular gland fibroblasts of tdTomato mice. The fluorescent signal from this cell line was observed in organs throughout the body, as well as in salivary glands. Primary culture cells derived from the submandibular gland were immortalized with SV40 large T antigen (GManSV cells); these cells exhibited increased migratory ability, as compared with those isolated from the sublingual gland. GManSV cells were tdTomato-positive and exhibited spindle-shaped fibroblastic morphology; they also robustly expressed mouse MSC markers: Stem cell antigen-1 (Sca-1), CD44, and CD90. This cell line retained multipotent stem cell characteristics, as evidenced by its ability to differentiate into both osteogenic and adipogenic lineages. These results indicate that Sca-1+/CD44+/CD90+-GManSV cells may be useful for kinetic studies of submandibular gland-derived MSCs in the context of in vitro co-culture with other types of salivary gland-derived cells. These cells may also be used for in vivo imaging studies, in order to identify novel cell therapy and regenerative medicine for the treatment of salivary gland diseases.