利用人類腎臟近曲小管表皮細胞建立三維細胞培養模型;Establish a three-dimensional in vitro model with human proximal tubule epithelial cell

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Title:	利用人類腎臟近曲小管表皮細胞建立三維細胞培養模型;Establish a three-dimensional in vitro model with human proximal tubule epithelial cell
Authors:	范毓婷;Fan, Yu-Ting
Contributors:	生命科學系
Keywords:	近曲小管表皮細胞;三維培養;腎毒性;體外模型;proximal tubule epithelial cell;3D culture;Nephrotoxicity;in vitro model
Date:	2021-01-27
Issue Date:	2021-03-18 16:51:18 (UTC+8)
Publisher:	國立中央大學
Abstract:	近年來在台灣接受血液透析及腎功能衰竭的患者數量迅速增加，除了主要的風險因素例如糖尿病和高血壓外，許多人還患有不明病因的慢性腎臟疾病（CKDu）。藥物濫用、飲食習慣或環境暴露（如食品安全問題和空氣、水污染問題），可能都是導致CKDu的潛在風險因素。目前對於確切肇成CKDu的了解有限，因此建立有效的腎細胞模型來預測其生物學反應和腎臟腎毒性至關重要。 HK-2細胞是來自人類腎臟近曲腎小管上皮細胞的永生化細胞株，表達許多轉運蛋白，其中多數與腎臟近曲小管對於藥物分泌至尿液或再吸收回血液有重大的相關性。因此HK-2細胞時常被用於腎毒性研究，然而二維細胞培養因環境過於單純，不易反應體內結構和微環境的複雜性。本研究利用臨床級明膠海綿作為支架來建立HK-2細胞的三維細胞培養模型。並進一步比較二維和三維細胞培養之間增殖的差異、表現形態和轉運蛋白等的基因表達，期為CKDu潛在因子建立一個有效的腎毒性測試平台。研究結果顯示在型態上三維培養的細胞呈現管狀分化，並比二維培養細胞表現較多的運輸蛋白。;In recent years, the number of patients receiving hemodialysis or even kidney failure has increased rapidly in Taiwan. In addition to predominant risk factors, such as diabetics and hypertension, many are suffered from chronic kidney disease of unknown etiology (CKDu). Drug-induced nephrotoxicity due to our medication consumption habit and environmental exposure such as worsen issues of food safety and air pollution can all be potential risk factors leading to CKDu. Given limited information about CKDu in Taiwan, it is crucial to establish an effective model to predict biological responses and renal damage to nephrotoxicity. HK-2 cell is an immortalized cell line from human renal proximal tubule epithelial cells, which expresses many transporters for renal drug secretion and urine compound reabsorption. The HK-2 cell line has been extensively used for nephrotoxicity study. However, the simplicity of the two-dimensional cell culture made it challenging to recapitulate the complexity of the in vivo architecture and microenviroment. Here, we developed a three-dimensional model for HK-2 cells using clinical grade-gelatin sponge as our scaffolds. We compared the cell viability, proliferation, morphology, and the gene expression of transporter markers between 2D and 3D culture. Our results showed that 3D cells displayed a tubule-like morphology and expressed more key transporters than 2D cells did. With this, we hope that we have established a more effective testing platform for nephrotoxicity and underline mechanism investigation of CKDu.
Appears in Collections:	[Graduate Institute of Life Science] Electronic Thesis & Dissertation

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