2012-08-012024-05-18https://scholars.lib.ntu.edu.tw/handle/123456789/704117摘要：源自懷孕時期子宮內胎體羊水之羊水幹細胞分化潛能為介於胚幹細胞與成體幹細胞時期之類多能性幹細胞，其表現Oct4 和 Nanog等胚幹細胞所表現之多能性標誌，但無胚幹細胞與誘導式多能性幹細胞所誘發之畸胎瘤產生之危險性，且其端粒長度亦顯著高於臍帶血與骨髓等來源之幹細胞，另因羊水幹細胞均源自於胎兒，未來應用於自體移植將不會有任何免疫排斥的問題。肝纖維化為慢性肝病，嚴重將導致肝硬化，且肝硬化為國人十大死因之一，因此建立肝纖維化之動物模式，以羊水幹細胞探討此疾病之幹細胞治療功效及相關機轉有其迫切性。然而，植入外源性幹細胞於體內是否直接進行分化、或是以細胞融合方式亦或是藉由胞外分泌因子來進行修補，迄今仍然具有爭議性與不確定性。因此，本三年期研究計畫，第一年擬以本實驗室已建立之螢光豬與螢光小鼠之羊水幹細胞，探討類肝臟細胞體外分化分析條件及肝纖維化小鼠幹細胞治療模式之建立。第二年將以表現紅色螢光蛋白質(enhanced red fluorescent protein)轉基因豬之羊水幹細胞(異種)，對於全身表現綠色螢光蛋白質之肝纖維化小鼠之治療模式及潛力，及綠色螢光蛋白質(enhanced green fluorescent protein) 轉基因小之鼠羊水幹細胞(同種異體)，對於全身表現紅色螢光肝纖維化小鼠模式動物之治療潛力。第三年將進一步探討此兩類表現螢光蛋白質細胞是否透過細胞融合來進行相關治療功效機轉之研究。綜言之，完成以上試驗將可提供羊水幹細胞進行臨床前幹細胞治療、基因治療或組織工程等極佳研究之用途及瞭解幹細胞治療功效之相關機轉。<br> Abstract: Amniotic fluid derived stem cells (AFSCs) which isolated from amniotic fluid of the pregnant female uterus, differentiation potentiality is between embryonic stem cell and somatic stem cell, can express Oct4 and Nanog pluripotent markers and without any teratoma formation risk that induced by embryonic stem cell and induced pluripotent stem cell (IPS). Especially, the telomere length of AFSCs is significantly higher than the bone marrow derived mesenchymal stem cell (BMMSC) and cord blood derived MSC. All of the amniotic stem cells were derived from fetus, as a result, when apply to the clinic in the future would not cause any immuno-rejection. Liver fibrosis is a chronic disease, occurred for a long term will lead to cirrhosis, is one of ten critical diseases causing mortality in Taiwan. Therefore, AFSCs will be used for cell source of stem cell therapy and simultaneously investigate whether they will ameliorate the liver fibrosis and related mechanism by mouse model. However, it still remains to be controversial and uncertain whether donor cells proceed to differentiate directly or cell fusion or paracrine mode to repair the lesion site. Hence, in this 3-year of research project, the first year will establish AFSCs derived from pig and mouse fetuses harboring with EGFP or ERFP are used to investigate the capability of hepatocyte-like differentiation and establish the mouse model of amniotic fluid stem cell therapy by liver fibrosis. The second year of this project will firstly effort to transplant xenogenic ERFP bearing pig AFSCs to EGFP bearing fibrosis mouse model and transplant allogenic EGFP bearing mouse AFSCs to ERFP bearing fibrosis mouse model to study the curing potential. The third year, we’ll investigate whether both EGFP and ERFP fluorescent proteins fusion or not to study the mechanism. In conclusion, our results will clarify related therapeutic potency and mechanism of liver fibrosis, and could be available in preclinical studies including AFSCs therapy, gene therapy and tissue engineering fields.綠色螢光基因轉殖小鼠紅色螢光基因轉殖豬羊水幹細胞肝纖維化EGFP (enhance green fluorescent protein)ERFP (enhance red fluorescent protein)Transgenic animalsAmniotic fluid stem cellLiver fibrosis