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題名:吳郭魚成魚心室肌細胞之初級培養     (18點)
Primary Culture of Ventricular Myocyte from Adult Tilapia
著者:童麗珠(Li-Chu Tung)
出版地區:台灣
出版城市:台北市
學科:生命科學
關鍵字:魚心室肌細胞 ; 酵素解離 ; 初級培養 ; 形態變化 ; Fish ventricular myocyte ; Enzyme dissociation ; Primary culture ; Morphological change
刊名:師大生物學報
卷期:30卷2期(1995.12)
頁碼:107-115
語言:繁體中文
摘要: 中文摘要PDF ; 英文摘要PDF

本研究主要在建立吳郭魚心室肌細胞的初級培養模式,並觀察細胞在培養過程中外形的變化。實驗方法是將剪碎的心室以protease-collagenase酵素溶液解離,大約可收獲40-50%的正常心肌細胞。將此心肌細胞置入EMEM培養液(內不含glutamine,再外加10%胎牛血青、一倍非必要基酸及抗生素)內,於維持在28℃的CO2培養箱內培養。剛分離的單一心室肌細胞呈細長桿形或長紡綞形-大小長約140-190μm、寬約3-5μm。在掃描電子顯微鏡下觀察,細胞是由數束肌原纖維(myofibrils)構成。細胞表面平滑,沒有橫管(transverse tubule)系統。剛開始培養的心肌細胞外形會漸變成圓形、橢圓形、短桿形或彎曲。培養至5-7天時,細胞才開始附著瓶底,附著率約1%。而後漸漸伸出長偽足向前後兩端延伸,並與鄰近的細胞相接合。此時的細胞會緩慢分裂增殖,甚至長成一片。細胞偶爾會收縮,常會在細胞片中央會形成一個收縮中心,啟動鄰近相連的細胞群同步收縮。在培養的末期,細胞會因收縮拉扯脫離瓶底,而退化死亡。在此環境下細胞並不能繼代及無限增殖,一般約可存活30-60天。
Ventricular myocytes of adult Tilapia were isolated enzymatically, cultured and examined their morphological changes at different time interval. Using mince digestion in protease and collagenase, cardiac muscle gave about 40-50% of isolated myocytes maintaining their spindle-shape. The size of the isolated myocytes were 140-190μm in length and 3-5μm in width. The culture medium was Eagle's minimal essential medium (MEM) with Earle's salts, supplemented with 10% fetal calf serum, 1 x nonessential amino acid mixture, 100 IUg/ml penicillin G, and 100μg/ml streptomycin. The cultured cells were grown at 28℃ in a humidified CO2 incubator. In order to eliminate fibroblast contamination, cells were cultured in the medium without glutamine. During the initial 24 hr culture, the cultured cells gradually shortened from their both ends and rounded up. Over 5 to 6 days postcultivation, the cells attached to the bottom of the culture flask, began to protrude pseudopodia and contact with each other. The cells proliferated slowly, and formed the cell sheet. The cells contracted spontaneously. The center of cell sheet often developed contractile center to synchronize the beating. The cells could not be subculture and also prolifeated indefinitely. The life span of cells in culture was 30 to 60 days.




本卷期目次
師大生物學報 30卷2期 (1995.12)
吳郭魚成魚心室肌細胞之初級培養/ 童麗珠
利用PCR與RFLP技術分析台灣產南方靈芝兩雜交不孕性群之遺傳變異/ 葉增勇張君玉李桂楨
瀕絕物種烏來杜鵑的族群內遺傳變異研究/ 黃生陳兆美許素玲呂勝由
哈盆溪臺灣馬口魚(Zacco barbata)之生殖生態/ 顏俊雄呂光洋陳永樹鄭振寬
補氣健脾藥四君子湯調節人體B淋巴球分泌A型免疫球蛋白/ 呂丹妮陳安德張玉冬曾哲明
碳源對Sordaria fimicola α-澱粉?釋出之誘導/ 李琦玫童武夫
 
   
 
   

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