大鼠、天竺鼠及豬的肺泡第二型上皮細胞分離與純化

Isolation and Purification of Alveolar Type Ⅱ Cells from Rats, Guinea Pigs, and Pigs

連毅(Yi Lien) ； 莊秀琪(Hso-Chi Chaung) ； 莊世宏(Shi-Hong Chuang) ； 鍾文彬(Wen-Bin Chung)

《中華民國獸醫學會雜誌》 26卷2期 (2000/06) Pp. 124-132

https://doi.org/10.7009/JTCSVS.200006.0124

摘要

肺泡第二型上皮細胞在肺臟主要可生成肺泡表面張力素、參與肺泡壁的修補，並具有生成趨化細胞素進而影響免疫反應及炎症反應的作用。本實驗根據前人分離大鼠肺泡第二型上皮細胞的方法加以修正，以簡易的方法且無需昂貴設備下，可同時分離高純度及存活率佳的豬隻肺泡巨噬細胞及肺泡第二型上皮細胞，並比較分離大鼠、天竺鼠及豬等三種動物的肺泡第二型上皮細胞的純度。所得的肺泡第二型上皮細胞並以polychrome染色法及電子顯微鏡分辨細胞的純度。大鼠、天竺鼠及豬分別採取10、10與5次重覆試驗後，得到以下的結果：大鼠純化之活細胞總數為9.7×10^6 cell/lung，肺泡第二型上皮細胞佔82.0±1.9％；天竺鼠活細胞總數為9.4×10^6 cells/lung，肺泡第二型上皮細胞佔83.7±2.5％；豬活細胞總數為1.62×10^7cells/lung，肺泡第二型上皮細胞佔81.2±1.5％。因此經修正後之分離方法可應用於較大型動物，且同時有效率的分離出純度高而存活率佳的肺泡第二型上皮細胞典與肺泡巨噬細胞。

關鍵字

大鼠 ； 天竺鼠 ； 豬 ； 肺泡第二型上皮細胞 ； 肺泡巨噬細胞

並列摘要

The alveolar type II cell performs a variety of significant vital essential functions within the lung, including surfactant metabolism regulation, alveolar repair in response to injury and chemokines secretion for inflammatory reaction modulation. This study improved the conventional means of isolating alveolar type II cells by digesting lung tissue with adequate amounts of elastase and trypsin, then employing the IgG ”panning” method to exclude similar sized immune cells as alveolar type II cells. This proposed method provides both a high yield and pure alveolar type II cell isolation. Alveolar type II cells were identified by the presence of characteristic lamellar inclusion bodies visualized by a polychrome stain and transmission electron microscopy AII of the viable cells per lung were 9.7x10^6 from the rats (n=10), 9.4x10^6 from the guinea pigs (n=10), and 16.2x10^6 from the pigs (n=5). The percentages of alveolar type II cells isolated from these three kinds of animals were 82.0±1.9%, 83.7±2.5%, and 81.2±1.5%, respectively. The results verify that this isolation procedure is feasible in large domestic animals since it causes little harm to the isolated cells.

並列關鍵字

Rat ； Guinea pig ； Pig ； Alveolar type Ⅱ cell ； Alveolar macrophage