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An attempt the tooth embryogenesis combined with embryonic stem cells and tooth mesenchymal stem cells that originated from neural crest

胚胎干细胞与神经嵴源性牙间充质干细胞联合牙胚发生的尝试

基本信息

批准号：
18592019
负责人：
YAMAKI Mariko
金额：
$ 2.35万
依托单位：
Matsumoto Dental University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2006
资助国家：
日本
起止时间：
2006 至 2007
项目状态：
已结题

项目摘要

We hypothesized that a combined use of ES cells and tooth mesenchymal cells is promising for successful development into tooth germ and/or teeth. Based on this hypothesis, we isolated mesenchymal cells from fetal mice, and established a stable cell line, called MDU1 line. We next in vitro cultured ES cells on MDU1 cells, which were used as feeder cells in the culture. Then we found that the ES cells developed into cells that produced keratin and the MDU1 cells into fibroblast-like cells that produced collagen. We therefore considered the cells thus co-cultured capable of developing into tooth germ and/or teeth. We also found that a mixture of cells co-cultured (ES and MDU1 cells) formed embryoid body (EB)-like spheres, called chimera Ebs. We next transplanted chimera Ebs with 3D scaffold made of type I collagen (called collagen sponge, CS) and transplanted the Ebs into sub-renal capsule in mice. Our close observations with histology and immuno-histochemistry identified extensive calcif … More ication, and significant development of odontoblasts and ameloblasts in the grafts. This identification indicates high potential of these Ebs as a good source for tooth germ and/or teeth, and therefore suggests that the absence of the entire structures is due to a small number of cells transplanted. Based on this suggestion, we plan to transplant a larger number of cells co-cultured to identify whether the cells develop into entire structures of tooth germ and/or teeth.We have observed during the course of the above experiments that ES cells did not form teratoma when transplanted with CS for 7 weeks, whereas ES cells readily formed teratoma when transplanted alone. This observation is extremely important because teratoma formation is one of the most critical problems in practicing stem cell therapy and no perspective for solving this problem has been reported so far. To obtain a better understanding of this phenomenon, we examined gene expression of ES cells, and identified expression of both undifferentiation genes and a tumor marker gene were significantly suppressed when the ES cells were co-cultured with CS. This suggests that CS can directly suppress teratoma formation, and also implies a good method to suppress teratoma formation from cells that experience specific induction from ES cells. We plan to examine this possibility as well. Less

我们假设ES细胞和牙齿间充质细胞的组合使用有希望成功发育成牙胚和/或牙齿。基于这一假设，我们从胎鼠中分离间充质细胞，建立了稳定的细胞系MDU 1。接下来，我们在MDU 1细胞上体外培养ES细胞，MDU 1细胞用作培养物中的饲养细胞。然后我们发现ES细胞发育成产生角蛋白的细胞，MDU 1细胞发育成产生胶原蛋白的成纤维细胞样细胞。因此，我们认为这样共培养的细胞能够发育成牙胚和/或牙齿。我们还发现，共培养的细胞混合物（ES和MDU 1细胞）形成了胚状体（EB）样球体，称为嵌合体EB。接下来，我们将嵌合体Ebs与由I型胶原制成的3D支架（称为胶原海绵，CS）一起移植，并将Ebs移植到小鼠的肾包膜下。我们通过组织学和免疫组织化学的密切观察发现， ...更多信息成牙本质细胞和成釉细胞在移植物中显著发育。这种鉴定表明这些EB作为牙胚和/或牙齿的良好来源的高潜力，因此表明整个结构的缺失是由于移植的细胞数量少。基于这一建议，我们计划移植更多的细胞共培养，以确定细胞是否发育成牙胚和/或牙齿的整个结构。在上述实验过程中，我们观察到ES细胞与CS一起移植7周时没有形成畸胎瘤，而ES细胞单独移植时容易形成畸胎瘤。这一观察结果是非常重要的，因为畸胎瘤形成是实践干细胞治疗中最关键的问题之一，迄今为止还没有报道解决这个问题的前景。为了更好地了解这种现象，我们检查了ES细胞的基因表达，并发现当ES细胞与CS共培养时，未分化基因和肿瘤标志物基因的表达均被显着抑制。这表明CS可以直接抑制畸胎瘤形成，并且还意味着抑制来自经历ES细胞特异性诱导的细胞的畸胎瘤形成的良好方法。我们也计划研究这种可能性。少

项目成果

期刊论文数量（0）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Prolonged in vitro culture and physical isolation of the inner cell mass increase the efficiency of human embryonic stem cell derivation

长时间的体外培养和内细胞团的物理分离提高了人胚胎干细胞衍生的效率

DOI：
发表时间：
2008
期刊：
Nature Methods (in press)
影响因子：
0
作者：
島津貴咲;高橋幸裕;矢島彩子;高島英造;古西清司;T.Yamashita;A.Chen
通讯作者：
A.Chen

XHAPLN3 is essential gene contributes to cardiogenesis during Xenous early development.

XHAPLN3 是 Xenous 早期发育过程中促进心脏发生的重要基因。

DOI：
发表时间：
2007
期刊：
影响因子：
0
作者：
島津貴咲;高橋幸裕;矢島彩子;高島英造;古西清司;T.Yamashita;A.Chen;T. Yamashita;A. Chen;T. Chan;T. Yamashita;A. Chen;T.Chan;C.Zhao;A.Hosoya;T.Ninomiya;C. Zhao;A. Hosoya;H. Nakamura;T. Ninomiya.;H. Nakamura.;Ozawa H.;Nakamura H.;Ozawa H.;Ozawa H.;Ozawa H.;Ozawa H.;小林泰浩;Y. Kobayashi;伊藤弓弦;Y. Ito;伊藤弓弦
通讯作者：
伊藤弓弦

Localization of perlecan and heparanase in Hertig's epithelial root sheath during root formation in mouse molars.

小鼠磨牙根形成过程中基底膜聚糖和乙酰肝素酶在赫蒂格上皮根鞘中的定位。