國立臺灣大學醫學院外科湯月碧2006-07-262018-07-112006-07-262018-07-112002-07-31http://ntur.lib.ntu.edu.tw//handle/246246/24471本計畫主要是應用基因轉殖於脂肪組織工程,在我們希望的部位自行產生脂 肪組織增生，以達到重建或整形的目的。過去我們在老鼠皮下注射基底膜 (basement membrane)的複合體Matrigel 或Cymetra 作為組織增生的支架，其中 混合可在生物體內分解的凝膠微粒，這些微粒為可控制的藥物釋放系統，可將我 們所使用的生長因子包在其中並逐漸釋放，以達到刺激細胞分化、增生的目的。 但此藥物釋放系統只在特定的時間內持續釋放生長因子,不可任意延長或中止。 而細胞經基因轉殖後不僅能穩定不斷地釋放我們想要的生長因子,且可經由調控 基因隨時中止或重新啟動其釋放,這也是基因療法的基本原理。基因療法已隨分 子生物學的突飛猛進日趨成熟,也是許多疾病治療希望所寄。我們在此將之應用 於自體脂肪組織再生工程,將必要生長因子的基因轉殖至脂肪幹細胞或其他間質 細胞如纖維母細胞等，使轉殖後之細胞能持續穩定地製造並釋放生長因子，我們 並在轉殖的基因上加上一段調控基因可隨時打開或關閉讓生長因子在必要時才 被製造及釋放。在基因轉殖載體方面,我們選擇正離子脂質體是因其具有轉殖效 率高、容易操作且實驗結果重覆性高等優點,並可避免病毒載體之危險性,是目前 廣被使用之基因轉殖載體。我們先在培養的細胞中評估,如果轉殖的效率、生長 因子的釋放與調控、甚至對細胞的毒性等皆有令人滿意的結果,我們將進一步做 動物實驗評估其將來應用於人體組織工程的可能性。The main purpose of this experiment is using gene transfection in the tissue engineering about adipogenesis. In the previous study, we used basement membrane complex (Matrigel or Cymetra) mixed with biodegradable gelatin microspheres, a controlled drug release system, was injected into subcutaneous layer of mice. bFGF, growth hormone and IGF-1 were incorporated into these microspheres respectively before mixing. The adipose precursor cells are fibroblast like precursor cel ls and could be stimulated by growth hormones to migrate into the scaffold and differentiate to mature fatty tissue. Although this drug releasing system could induce adipogenesis, the growth factors were released continuously but could not be stopped or restarted releasing at any time as we will. If we transfect genes of growth factors we need with regulatory genes into adipocyte precursor cells or other mesenchymal cells such as fibroblasts, the desired growth factors could be released stably and the genes could be turned on or off at any time we need. The cationic lipid reagents are the transfection system of our choice because it yields high transfection efficiency in a wide variety of eukaryotic cells, is simple to perform, and ensures consistently reproducible results. Here in vitro studies by transfection of desired genes into cultured cells were tried, if the results are satisfactory, we will apply it further to in vivo studies.application/pdf165059 bytesapplication/pdfzh-TW國立臺灣大學醫學院外科組織工程脂肪組織增生生長因子基因轉殖調控基因正離子脂質體reporthttp://ntur.lib.ntu.edu.tw/bitstream/246246/24471/1/902314B002414.pdf