靜水壓對細胞分化及增生影響的實驗研究

Experimental study of hydrostatic pressure effects on cell differentiation and growth

余明翰(Ming-Han Yu)

指導教授 ： 鍾志昂

國立中央大學/工學院/機械工程學系/碩士(2012年)

摘要

組織工程為一個隨著現今醫療技術發展而新興的組織修補方法，目標為發展出由體外培養的方式自行培養所需的組織，經手術移植進入患者體內更換受損的組織，而為了發展體外培養需藉助生物反應器的幫助，生物反應器除提供適合細胞生存的環境之外，更可以提供不同的機械刺激調控細胞的行為或是基因表現等。
本文旨在探討細胞在靜水壓刺激下的分化以及增生行為。因此設計出一套低壓力且壓力精度高之生物反應器，為得到穩定且準確的壓力值，使用回授控制系統，此外使用Labview軟體進行人機介面的建立。建立的生物反應器可以施加定壓或循環壓力，定壓下壓力值最大可以達到150 kPa，最小值為10 kPa，而循環壓力頻率可以達到 1 Hz，系統控制精度為 1.5 kPa。
在細胞分化實驗部份我們使用此生物反應器給予人類胎盤源多功能幹細胞 四種不同定常壓力刺激 (0 kPa、10 kPa、30 kPa、50 kPa )及在細胞中添加Osteogenic (Ost) 的藥物刺激誘導幹細胞朝向成骨細胞分化。用Alizarin Red Stain (ARS) 檢測壓力刺激對細胞分化的結果。而抑制細胞增生實驗部份，我們對於膀胱癌症細胞進行三種不同的定常壓力測試(0 kPa、10 kPa、100 kPa) ，再用Methylthiazolyldiphenyl-tetrazolium bromide分析樣本上存在的細胞數量。結果顯示當定常壓力越大的時候，幹細胞分化的程度越好，且抑制癌症細胞增生的表現也越好。

關鍵字

生物反應器 ； 靜水壓 ； 細胞 ； 分化 ； 增生

並列摘要

Tissue engineering is a novel way to repair the injured tissue. Tissue engineering aims to culture biologically functional tissue in vitro, that can be transplanted to replace damaged tissue. In order to develop in vitro culture, the bioreactor plays an important role. The bioreactor provides a suitable environment for cell survival, and apply different mechanical stimulation.
This study investigats the cell behaviors of differentiation and proliferation in hydrostatic pressure stimulation. A bioreactor with low-pressure and high accuracy is bulit. For achieving the stable and accurate pressure values, we used a feedback control system and set up the man-machine interface by the software Labview. The system can generate constant pressure and the periodic pressure. The maximum constant pressure is 150 kPa and the minimum is 10 kPa. The frequency of periodic pressure can reach 1 Hz. The pressure precision is about 1.5 kPa. We used the bioreactor to culture Human Placenta-Derived Multipotent Cells (PDMCs). We gave four different constant pressures (0 kPa,10 kPa, 30 kPa and 50 kPa) and added the Osteogenic (Ost) to induce the stem cell toward the osteoblast differentiation. Differentiation levels were assessed by Alizarin Red Stain (ARS). In the experiments of cell proliferation, we gave three different constant pressures (0 kPa, 10 kPa and 100 kPa) to Human bladder papillary transitional cell carcinoma. The number of cells is calculated by Methylthiazolyldiphenyl-tetrazolium bromide assay. Results of the experiments showed that applying constant pressure was able to promote the differentiation of PDMCs and reduce the proliferation of the carcinoma cells.

並列關鍵字

bioreactor ； cell ； proliferation ； hydrostatic pressure ； differentiation

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