2014-10-012024-05-18https://scholars.lib.ntu.edu.tw/handle/123456789/707519摘要：本子計畫的內容是將生醫高分子材料應用於幹細胞純化篩選，並進行生醫材料上自組裝幹細胞球 之轉譯研究。在前一期的計畫中，我們發現間葉幹細胞在特定高分子材料上培養，能夠自組裝形成三 度空間的幹細胞球體，而有助於維持或提昇多潛能基因(Nanog,Oct4, Sox2)的表現，並使細胞具有較高 的分化潛能；此外我們也發現在細胞自組裝的過程中可將基因轉殖進入細胞。本期計畫有三個主要標 的：(1)利用生醫高分子材料調控細胞的骨架蛋白、貼附與聚集等行為，分離純化出特定的間葉幹細胞 族群，尤其是含CD271的細胞比例，並建立材料篩選幹細胞相關的分子機制。(2)利用生醫材料篩選與 自組裝之幹細胞球體作為細胞來源，結合生醫材料支架應用於動物關節軟骨缺損之再生轉譯，並評估 不同材料上自組裝之幹細胞球體在修復效果的差異；同時在球體形成時轉殖有助於心肌再生的因子(如 GATA4)，提供給總計畫的其餘子計畫進行動物心肌修復的評估。(3)設計高分子材料以探討幹細胞自 組裝之行為，藉由合成一系列不同化學組成、軟硬度或親疏水性等之材料，確認造成幹細胞形成球體 之關鍵材料因素。透過此計畫，期望能利用高分子材料來分離與純化特定幹細胞族群，並能夠經由轉 譯在未來應用於臨床幹細胞治療。<br> Abstract: Stem cells have the ability to self-renew and maintain their undifferentiated state, and may differentiate upon appropriate physical and chemical inductions. They are, however, susceptible to loss of stemness in an in vitro culture environment. In the period of last project (2011~2014), we have discovered that the self-renewal of mesenchymal stem cells (MSCs) was well maintained on a few selective biomaterials by forming three-dimensional (3D) cellular spheroids. Plasmid DNA may be transferred into MSCs during spheroid formation. In the new sub-project (2014~2017), we will further use biomaterials to select specific population of MSCs and translate the research of MSC spheroids in animal studies. Therefore, in this sub-project, we aim at: (1) establishment of new selection methods and identification of the mechanism for biomaterial substrate-based cell selection; (2) translating the studies of substrate-derived MSC spheroids; and (3) designing model substrates. First, we will establish the protocols to maximize the potential markers (in particular CD271) through biomaterial selection. We will also analyze the integrin and cadherin expressions to explore the mechanism for cell selection. Second, the selected MSCs will be loaded in 3D printed scaffolds and implanted in the osteochondral defects of rabbit knees to verify their therapeutic potential of the biomaterial-selected stem cells. We will also evaluate the efficacy of various substrate-derived MSC spheroids in the osteoarthritis rabbit model for cartilage repair. Besides, we will use biomaterials to deliver GATA4 plasmid to MSCs, and verify the effect of genetically-modified MSC spheroids in porcine heart regeneration (collaboration with another sub-project of main project). Third, we will synthesize the biomimetic materials that promote the self-assembly of MSC spheroids and identify the critical properties of a substratum material for promoting the spheroid formation and gene delivery. Through the execution of this sub-project, we expect that materials technology may be expanded to isolate cells, select cells, as well as prepare cells for clinical stem cell therapy in the future.間葉幹細胞細胞篩選基因傳遞軟骨再生組織工程mesenchymal stem cells (MSCs)cell selectiongene deliverychondrogenesistissue