牙胚间充质与干细胞参与构建的再生牙大小的相关性研究
发布时间：2019-03-02 17:19
【摘要】：诱导多能干细胞(iPS)技术作为一种新的医疗策略,在牙齿修复领域有广阔的应用前景。多种来源的细胞已被证实可以转化为iPS细胞。同时,这些iPS细胞兼具有组织记忆性,使得之后的诱导过程更为简易有效。课题组先前的实验已成功将人源iPS细胞诱导分化为具有成牙潜能的上皮细胞,在与胎鼠口腔间充质共培养之后移植到裸鼠中成功再生出牙齿。为了进一步发现该牙齿再生模型的可行性以及间充质对牙齿再生的影响,我们进行了本课题的研究。本研究将人尿液来源的诱导多能干细胞诱导分化为上皮样细胞,利用qPCR以及免疫荧光检测其最适合的移植时期,从而选得上皮细胞。实验将E48天的胎猪中的牙胚进行消化分离得到口腔间充质,将该间充质以及消化上皮样细胞得到的上皮细胞层在体外重新结合后共培养1-2天移植入裸鼠肾包膜上进行牙齿的再生。之后对比先前实验中鼠间充质来源的再生牙齿和猪间充质来源的再生牙齿的大小以及承压率,并对牙齿进行HE染色等实验。结果显示,通过对上皮细胞相关基因进行检测发现人尿来源的iPS细胞诱导的上皮细胞层在第7天的时同间充质的结合能力最为优异。将该上皮细胞层同胎猪口腔间充质重新结合后共培养的组织块移植入肾包膜后成功再生出牙齿。统计发现猪牙胚再生的牙齿的成牙率为92.86%,猪间充质共培养再生牙齿的成牙率为72.97%,远高于已发表文章中鼠间充质共培养再生牙齿(21.81%)的成牙率。同时,所有生成的类牙齿组织都大于鼠口腔间充质来源的。实验中移植的牙齿在20周时,其上皮细胞分化为成釉细胞,并伴有牙髓的生成。综上所述,实验表明,我们的人iPS再生牙齿模型有很好的适用性,同时口腔间充质对牙齿的再生有很大的影响。这为我们之后使用人源iPS细胞再生出牙齿提供了良好的基础,加深了我们对口腔间充质的认知,为之后人的牙齿再生应用起一定的推动作用。
[Abstract]:As a new medical strategy, induced pluripotent stem cells (iPS) has broad application prospects in dental restoration. Cells from a variety of sources have been proved to be able to transform into iPS cells. At the same time, these iPS cells also have tissue memory, which makes the induction process easier and more effective. Previous experiments in our team have successfully induced human iPS cells to differentiate into epithelial cells with odontogenic potential, and successfully regenerated teeth in nude mice after co-culture with fetal rat oral mesenchymal cells. In order to find the feasibility of the tooth regeneration model and the effect of mesenchymal on tooth regeneration, we carried out this study. In this study, human urine-derived induced pluripotent stem cells were induced to differentiate into epithelial-like cells. QPCR and immunofluorescence were used to detect the most suitable period of transplantation to select epithelial cells. Oral mesenchymal tissue was obtained by digesting and separating tooth germs from E48-day-old pigs. The epithelial cell layer obtained from the mesenchymal and digestive epithelial-like cells was recombined in vitro and then co-cultured for 1 day and 2 days after transplantation into the renal capsule of nude mice for tooth regeneration. Then we compared the size and compression rate of the regenerated rat mesenchymal teeth and porcine mesenchymal regenerated teeth in previous experiments, and the teeth were stained with HE. The results showed that the epithelial cell layer induced by human urine-derived iPS cells had the best ability to bind to mesenchymal cells on the 7th day after the detection of epithelial cell-related genes. The epithelial cell layer was recombined with fetal porcine oral mesenchymal tissue and the co-cultured tissue was transplanted into the renal capsule to regenerate the teeth successfully. It was found that the rate of tooth formation was 92.86% for porcine tooth germ regeneration and 72.97% for porcine mesenchymal co-culture regeneration teeth, which was much higher than that of mouse mesenchymal co-culture regeneration teeth (21.81%) in published articles. At the same time, all the generated tooth-like tissues were larger than those derived from mouse oral mesenchymal tissue. At 20 weeks after transplantation, the epithelial cells of the transplanted teeth differentiated into ameloblasts, accompanied by pulp formation. In conclusion, the experiment shows that our human iPS dental regeneration model has good applicability, and oral mesenchymal has a great effect on tooth regeneration. This provides a good basis for us to regenerate teeth by using human iPS cells, deepen our understanding of oral mesenchymal tissue, and promote the application of human tooth regeneration.
【学位授予单位】：吉林大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R78;R318.08
本文编号：2433278
[Abstract]:As a new medical strategy, induced pluripotent stem cells (iPS) has broad application prospects in dental restoration. Cells from a variety of sources have been proved to be able to transform into iPS cells. At the same time, these iPS cells also have tissue memory, which makes the induction process easier and more effective. Previous experiments in our team have successfully induced human iPS cells to differentiate into epithelial cells with odontogenic potential, and successfully regenerated teeth in nude mice after co-culture with fetal rat oral mesenchymal cells. In order to find the feasibility of the tooth regeneration model and the effect of mesenchymal on tooth regeneration, we carried out this study. In this study, human urine-derived induced pluripotent stem cells were induced to differentiate into epithelial-like cells. QPCR and immunofluorescence were used to detect the most suitable period of transplantation to select epithelial cells. Oral mesenchymal tissue was obtained by digesting and separating tooth germs from E48-day-old pigs. The epithelial cell layer obtained from the mesenchymal and digestive epithelial-like cells was recombined in vitro and then co-cultured for 1 day and 2 days after transplantation into the renal capsule of nude mice for tooth regeneration. Then we compared the size and compression rate of the regenerated rat mesenchymal teeth and porcine mesenchymal regenerated teeth in previous experiments, and the teeth were stained with HE. The results showed that the epithelial cell layer induced by human urine-derived iPS cells had the best ability to bind to mesenchymal cells on the 7th day after the detection of epithelial cell-related genes. The epithelial cell layer was recombined with fetal porcine oral mesenchymal tissue and the co-cultured tissue was transplanted into the renal capsule to regenerate the teeth successfully. It was found that the rate of tooth formation was 92.86% for porcine tooth germ regeneration and 72.97% for porcine mesenchymal co-culture regeneration teeth, which was much higher than that of mouse mesenchymal co-culture regeneration teeth (21.81%) in published articles. At the same time, all the generated tooth-like tissues were larger than those derived from mouse oral mesenchymal tissue. At 20 weeks after transplantation, the epithelial cells of the transplanted teeth differentiated into ameloblasts, accompanied by pulp formation. In conclusion, the experiment shows that our human iPS dental regeneration model has good applicability, and oral mesenchymal has a great effect on tooth regeneration. This provides a good basis for us to regenerate teeth by using human iPS cells, deepen our understanding of oral mesenchymal tissue, and promote the application of human tooth regeneration.
【学位授予单位】：吉林大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R78;R318.08
【参考文献】
相关期刊论文 前1条
1 ;Making a tooth:growth factors,transcription factors,and stem cells[J];Cell Research;2005年05期
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