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題 名 | Culture of Retinal Pigment Epithelium from Evisceration Specimens=採用眼球剜除術後標本所培養出之視網膜色素上皮細胞 |
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作 者 | 曾漢儀; 吳文權; 高英賢; 吳弘鈞; | 書刊名 | The Kaohsiung Journal of Medical Sciences |
卷 期 | 20:5 2004.05[民93.05] |
頁 次 | 頁225-229 |
分類號 | 416.746 |
關鍵詞 | 視網膜色素上皮細胞; 組織體外培養; 酵素分離法; Retinal pigment epithelial cells; Explantation; Enzymatic dissociation; |
語 文 | 英文(English) |
中文摘要 | 過去尚未有以眼球剜除術後標本試做人類視網膜色素上皮細胞分離和培養之報告。本研究利用兩個嚴重眼球外傷的個案取其眼球剜除之標本進行視網膜色素上皮細胞之培養。我們採用兩種不同的細胞分離方法。個案一採傳統組織體外培養法,結果發現雖然視網膜色素上皮細胞順利生長,但是被纖維母細胞,黑色素細胞等其他種類細胞污染而駁雜不純。個案二採用酵素分離法,先使用 0.25% 胰蛋白脢將視網膜色素上皮細胞自脈絡膜取下再移至培養皿即可成功抑制其他細胞之污染。得到純種視網膜色素上皮細胞之後採用 F12 培養基加入 30% 胎牛血清可將視網膜色素上皮細胞繼續繁衍至五個世代。此視網膜色素上皮細胞經採用免疫細胞染色法證實確含有上皮細胞特有之細胞角質素無誤。本研究中的細胞分離及培養方法提供了人類視網膜色素上皮細胞培養的另一選擇,並且可對視網膜色素上皮細胞的細胞生物學及病理生理學研究有所裨益。 |
英文摘要 | Human retinal pigment epithelial (RPE) cell cultures are usually obtained from donor eyes; isolation and culture of RPE cells obtained by evisceration has not been reported previously. The present study attempted to isolate and cultivate RPE cells from evisceration specimens obtained from two cases with severe ocular trauma. Two different isolation methods, explantation and enzymatic dissociation, were used. In Case 1, RPE cells grew from the explants, but were contaminated with other cells such as fibroblasts and melanocytes, and no pure RPE cultures were obtained by explantation. In Case 2, RPE cells were separated from choroids using 0.25% trypsin before plating for culture, which effectively eliminated contaminating cells. A pure RPE culture was obtained and cultured with F12 medium supplemented with 30% fetal bovine serum. With this enzymatic dissociation method, cultured RPE cells grew to confluence in primary culture and could be maintained in culture for five passages. Cultured RPE cells were identified by the presence of cytokeratin expression, as shown by immunocytochemical staining. These isolation and culture methods provide alternative sources for human RPE cells and could be useful in studies of the cell biology and pathophysiology of human RPE cells. |
本系統中英文摘要資訊取自各篇刊載內容。