• 作者： 劉杭生; 邴俊棠; 楊永孚; 趙壯飛
• 作者服務機構： 三軍總醫院婦產部; 國防醫學院生物及解剖學科; 三軍總醫院臨床醫學研究部細胞生物研究室
• 中文摘要： 本實驗中，吾人利用3T3細胞株建立之方法，由14天大小之純種金黃色倉鼠胚胎建立出此GHEF-Ⅲ正常纖維芽細胞株。經稀釋篩選後，已再殖超過一年之久；此細胞在培養下表現出單層生長，並且有接觸抑制之特性。在光學顯微鏡、穿透式顯微鏡及免疫螢光檢查觀察下，此梭形細胞有一大而橢圓的胞核，胞質內富含顆粒內質網及粒線體，其細胞骨架vimentin在胞質內呈現散亂網狀分佈並在胞核處形成帽蓋，但在胞質內卻無cytokeratin之存在。此結果顯示GHEF-Ⅲ細胞的來源是纖維細胞。CHEF-Ⅲ細胞在體外培養下，並無黴漿菌或細菌之感染，其倍殖期為15小時，染色體分析顯示其以雙套44個之形成存在。由於GHEF一III細胞具有接觸抑制，無法在軟瓊脂培養基中生成細胞聚落，及在裸鼠身上無法生成腫瘤之特性，故顯示此細胞未發生轉形，仍具有正常細胞之特性。當CHEF-Ⅲ細胞以人類膀胱癌致癌基因質體pT-C從事轉染實驗時，可見此細胞能被pT-C致癌基因產生以下轉形有喪失接觸抑制，細胞骨變形，可在低血清濃度下生長，染色體異常分佈，獨立停泊及在裸鼠身上生腫瘤。由上述GHEF-Ⅲ細胞在體外培養下之轉形試驗結果，可提供吾人對致癌基因在致癌過程中所扮演之角色有所了解。且由於細胞具有穩定之基因結構及較少之自發性變形，吾人相信此建立之倉鼠胚胎纖維芽細胞株，為一理想的可轉染DNA之接受細胞。
• 英文摘要： A diploid, continuous cell line, Golden Hamster Embryo Fibroblast-Ⅲ (GHEF-III), which hadbeen passaged for one year, was established essentially by a 3T3 protocol from primary culture of 14-day-gestation Golden Hamster embryo fibroblast cells. The cultured fibroblast exhibited monolayer growth andhad contact inhibition. In morphological identification by light microscope (LM), transmission electronmicroscope (TEM) and immunofluorescence examination (IF), these multipglar and spindle-shaped cellshad a large ovoid nucleus, enriched rER and mitochondria in the cytoplasm. On the other hand, thevimentin presented in the cell with a random network and capped around the nucleus. The results in-dicated that the cultured GHEF-III cells were fibroblast in origin. The cells were free of bacterial andmycoplasma contamination. The doubling time in GHEF-Ⅲ was about 15 hours. Chromosomal analysisof GHEF-Ⅲ presented a diploid stem cell line with a modal number of 44. No evidence of transformationof GHEF-Ⅲ was shown by properties of contact inhibition, no colony formation in soft agar, and notumor growth in nude mice. The transformation of GHEF-Ⅲ after transfection with pT24-C3, an oncogenicplasmid, was shown by the evidence of loss of contact inhibition, growth in low serum medium, colonyformation in soft agar and tumor growth in nude mice. In vitro transformation testing of these cells mayprovide valuable data in studying the role of tumor transforming genes in carcinogenesis. Owing to thegenetic stability and less spontaneous transformation, this GHEF-Ⅲ cell line can be utilized as a source ofrecipient cells in transfection assay.
• 中文關鍵字： transfection; oncogene; transformation.
• 英文關鍵字： --