• 作者： 趙壯飛; 趙子健; 呂美華; 盧昭元; 陳幸一
• 作者服務機構： 國防醫學院生物及解剖學科暨三軍總醫院醫學研究部; 國防醫學院醫學科學研究所; 國防醫學院生理及生物物理學科暨慈濟醫學研究中心
• 中文摘要： 本實驗以“組織塊種植法”成功地由原發性高血壓大白鼠及正常血壓大白鼠之主動脈培養出高純度的內皮細胞。首先，將血管塊置於以纖維網蛋白處理過之培養皿上，然後再分別測試不同條件下對內皮細胞生長的影響，包括：處理血管時氧的供給、植種血管塊時培養皿的傾斜及血管塊移除的時間等，結果顯示這些條件顯然對細胞的生存、組識之貼附及減少非內皮細胞之污染等非常重要。培養之細胞再以形態上、免疫組織化學上及生化上之方法來定性時顯示這些細胞對von Willerbrand因子之抗體呈正反應，且可吞噬D1I-ac-LDL，可見其具內皮細胞細胞之特徵。同時，這些內皮細胞分泌前列腺素I2、血栓素及內皮素之能力也被測試，結果顯示培養之自發性高血壓大白鼠內皮細胞分泌前列腺素I2及內皮素之能力比正常血壓大白鼠之內皮細胞高，但其分泌血栓素之能力則相同。因此，以“組織塊種植法”可在體外成功地培養出具生物功能之高純度內皮細胞，利用這個方法可使我們更進一步的研究內皮細胞在高血壓形成過程中所扮演的角色。
• 英文摘要： In the present study, we successfully established a "tissue explant technique" to obtain high yieldand purity of endothelial cells from the aorta of hypertensive and normotensive rats (SHR and WKY).Small pieces of aorta were placed on fibronectin precoated petri dishes. The effects of oxygenation in thetissue preparation stage, tihing of the petri dish during the explanting period and timing of the removalof tissue blocks from petri dishes were evaluated. These procedures appeared to be critical for cell survival,tissue adhesion and minimizing of non-endothelial cell contamination. The cultured endothelial cells werecharacterized by morphological, immunohistochemical and biochemical examination. The cultured cellsfrom both SHR and WKY rats showed similar endothelial cell character, positive immunofluorescencestaining for the von Willebrand factor, and uptake of acetylated low-density lipoprotein (DiI-ac-LDL).The secretory function of prostacylcin I2 (PGI2), thromboxane A2 and endothelin of cultured endothelialcells was measured. The results showed that the secretion of both PGI2 and endothelin was greater inSHR than in WKY rats, but that there was no difference in thromboxane A2 secretion. Therefore, our"tissue explant technique" can provide high yield and purity of endothelial cells with their specific biologicalfunction in vitro. It will permit us to further study the role of endothelial cells in the development ofhypertension.
• 中文關鍵字： rat; culture; aortic endothelial cells.
• 英文關鍵字： --