- 作者: 陳華宗; 陳家福; 呂福江; 鄒順生; 張子明; 孫綿方
- 作者服務機構: 國防醫學院暨三軍總醫院外科部一般外科; 國防醫學院臨床醫學部; 國防醫學院病理部; 加拿大多倫多大學生理系
- 中文摘要:
十八雜種狗胰臟用來進行狗胰島分離研究。分離方法,經胰灌注結締組織?使胰臟漲大,利用自動篩檢法使胰島從
胰臟組織中分離出來,再利用Histopaque Ficoll密度差將胰島和胰腺體組織分開並純化狗胰島。分離狗胰島的活性功
能鑑定,體外實驗採用組織學及胰島灌注法來評估;體內實驗則利用糖尿病裸鼠異種移植模型來評估其功能。
結果:有十二次實驗分離純化結果良好。純化前後的胰島數目分別為4767.1±560.1及3637.7±333.4等積胰島/每克
胰組織。其純度高於百分之九十。組織學檢查,經Aldehyde Fuchsin染色顯示胰島中含有大量β顆粒。胰島灌注結
果,胰島經16.7mM高濃度葡萄醣及1mM IBMX刺激,其胰島分泌量是1.67mM葡萄醣刺激分鉍量的12.93±4.75倍。十
隻糖尿病裸鼠接受狗胰島移植後,血醣維持正常達兩週有七隻,達三週有五隻。
結論,利用自動篩檢法及Histopaque Ficoll密度差可提供大量良好高純度的狗胰島,其活性功能經體外及體內實
驗研究證實完好。此可援引為人胰島分離研究的理想動物模式。
a - 英文摘要: Eighteen pancreata from adult mongrel dogs were used for the study of islet isolation. The pancreaswas distended with collagenase in Hanks' solution. The automated screen method and Histopaque Ficollgradients were used to isolate and purify the canine islets. In vitro, the viability of isolated islets was as-sessed by both histology and perifusion studies. In vivo, the islet function was evaluated by using a nudemice xenograft model. Fair to good isolation and purification was found in 12 experiments. Before and after purification,the isolated islet count was 4767.1±560.1 and 3637.7±333.4 islet equivalence (I.E.)/gm pancreatic tissue.The purity was above 90%. Aldehyde Fuchsin stain disclosed islets with copious beta granules. The stimula-tion index of islets responding to 16.7 mM glucose plus 1 mM 3-isobutyl-l-methylxanthine (IBMX) versus1.67 mM glucose was 12.93±4.75. Normoglycemia was restored and maintained for up to 2 weeks in 7of 10 and up to 3 weeks in 5 of 10 diabetic nude mice transplanted with canine islets. In conclusion, the automated screen method and Histopaque Ficoll gradients afford a good yieldof highly purified canine islets, and functional viability was verified both in vitro and in vivo. This willbe an ideal model for isolation of human islets.
- 中文關鍵字: islets of Langerhans; isolation; purification; transplantation
- 英文關鍵字: --