The application of Antrodia camphorata extracts used for maintaining stem cell pluripotency

• Main Authors: Yi-Hsuan Lee, 李怡萱
• Other Authors: Shih-Ping Liu
• Format: Others
• Language: zh-TW
• Published: 2013
• Online Access: http://ndltd.ncl.edu.tw/handle/13660456829511291505

碩士 === 中國醫藥大學 === 基礎醫學研究所碩士班 === 101 === 　　Embryonic stem (ES) cells are pluripotent stem cells, that have the ability to differentiate into all types of cells. For this reason, ES cells have the potential to clinical application. Cultivation of the ES cells need leukemia inhibitory factor (LIF) to maintain ES cells self-renewal by activating Jak2/Stat3 signaling pathway. However, LIF is an expensive reagent. The goal in this study is to find out a traditional Chinese medicine extract which can replace LIF to maintain the pluripotency of ES cells. In our previous study, we found that Ethanol Extracts Antrodia Camphorata (EEAC) could increase the gene expression leves of Oct4 and Sox2, the genes that could maintain the stemness of stem cells. For this reason, AC has the potential to replace LIF in ES cells cultivation. ES cells were treated with different concentrations of EEAC and identified the stemness of ES cells by Alkaline phosphatease (AP) and immunofluorescent staining. We observed these cells expressed the characteristic of ES cells, including AP, Nanog, and SSEA1 at the concentration of 0.8 μg ml-1. Furthermore, the ES cells were passaged for six generation by the culture medium containing EEAC 0.8 μg ml-1, then used embryoid body formation to identify the pluripotency of ES cells. The results showed that the ES cells still could differentiate to the three germ layer including Tuj1, α-SMA, and Gata4. Finally, we wanted to know why EEAC could maintain the stem cell pluripotency and find the major pathway. By the data of western blot, Q-PCR and ELISA, EEAC activated Jak2/Stat3 pathway and increased the gene expression of related cytokines resulting in maintaining the ES cells self-renewal and pluripotency. In summary, we demonstrated that EEAC could maintain ES pluripotency by activing Jak2/Stat3 signaling pathway.