利用微流體晶片進行單一樹突狀細胞與癌細胞之電融合以生產高效率的癌症疫苗

The on-chip high- efficient electrofusion of dendritic cells and tumor cells for efficient cancer vaccine generation

呂仲桓(Chung-Huan Lu)

指導教授 ： 劉承賢

國立清華大學/工學院/動力機械工程學系/碩士(2014年)

摘要

由於人們生活型態的轉變，癌症在國內已位於十大死因之首長達三十一年，如今每一百人就有二十八人死於癌症。惡性腫瘤是癌症的病原，其形成原因大多數為先天性突變、致癌基因的過度表現。癌細胞具有強大的增生能力及轉移轉化能力，且由於致癌基因的活化需長時間的突變，因此大多要長時間後才能被診斷出，所以癌症在早期被視為一種絕症。然而隨著醫療的進步，切除手術、化療等相繼被應用於治療惡性腫瘤。切除手術雖然能直接將腫瘤切除，但若癌細胞已轉移到其他部位，此方法只能暫緩病情；化療雖然能較有效的運用藥物治療癌症，但其副作用非常大，常令患者為之卻步。雖然早期已有上述應對癌症的方式，但它們都無法有效地根治，因此近期有許多種新興的療法相繼提出，如基因療法、抑制血管新生、抑制促癌細胞蛋白質及免疫療法等。
免疫療法是多種治療方式中最具潛力的選項。2010年4月29日FDA批准了Provenge癌症治療疫苗。Provenge用於晚期前列腺癌的治療，也代表了客製化醫療新時代的來臨，人們可以根據不同種類的癌症用不同的疫苗做治療。在眾多的癌症免疫療法策略中樹突狀細胞/腫瘤融合治療疫苗是一門最受矚目選項。樹突狀細胞是吞噬細胞的一種，其具有最強的抗原表現，當樹突狀細胞吞噬抗原後，會活化適應性免疫系統使身體產生防衛的效果並有效的治療癌症。因此，將樹突狀細胞與分裂快的癌細胞融合，即可大量製作疫苗。
但過去使用具二乙醇化學電融合或隨機性電融合製造出的樹突狀融合疫苗，雖可誘導毒殺性 T 淋巴球的活化，但其融合效率以及品質十分不穩定。因此在本論文希望透過生物微機電利用半導體製程，在細胞融合的製作技術上做改良。製作在微米尺度的結構下完成精確且大量的細胞配對之生物晶片，配合輔助電極，在10伏特以下的電壓進行細胞膜電穿孔，做高良率的細胞電融合克服並改善目前細胞融合技術的關鍵障礙，使其能簡易的完成大量、高良率的癌症免疫疫苗。

關鍵字

細胞電融合 ； 微流體晶片 ； 細胞配對 ； 樹突細胞 ； 腫瘤細胞 ； 癌症疫苗

並列摘要

Because people lifestyle changed, cancer has been the top ten causes of death for 13 years. In every 100 people, 28 people were dead because of cancer. Cancer is caused by either the congenital mutations or the overexpression of oncogenes. It has a powerful ability to transform proliferative and transfer. Because the activation of oncogenes mutations takes months, the diagnosis need a long time. Therefore, the cancer was considered a terminal illness. However, with the medical improvement, the surgery and chemotherapy have been used to cure malignant tumors. Although surgery is easy to remove the tumor directly, this method can only postpone the disease if the cancer has moved to other organs. The chemotherapy can effective cure the cancer, but the patients might usually suffer from the significant side effects. However, both of these methods are unable to effectively cure the cancer. There are many kinds of therapies proposed in recent years, such as gene therapy, inhibition of angiogenesis, inhibition of pro-cancer protein and immunotherapy.
Immunotherapy has the potential among a variety of treatments. Cancer immunotherapy is a popular medical technology. On April 29, 2010, the FDA approved cancer treatment vaccine which was called Provenge. Provenge is used for the treatment of advanced prostate cancer. It represents the customization of new medical era. It can be used to make different vaccines according to different types of cancer. Among all strategies, cancer immunotherapy of dendritic cells / tumor fusion vaccine is a therapeutic option. Dendritic cells are phagocytes and have the strongest antigen expression. When dendritic cells swallow the antigen, they will activate the adaptive immune system. Therefore, the fusion of dendritic cells / tumor can make dendritic cells divide faster and mass-produce the vaccine.
In tradition, either polyethylenglycol or randomness method was used for electric fusion. The efficiency and the quality of the vaccine were both unstable. Through this research we develop a cell electrofusion lab chip which combined the rapid and precise cell pairing micro-structures and the high yield electrofusion micro-electrodes to improve the cell fusion techniques and overcome the key barriers. We wish to build and develop an automated, mass-produced and efficient dendritic cell / tumor fusion vaccine.

並列關鍵字

Cell electrofusion ； Microfludics chip ； Cell trapping ； dendritic cell ； tumor cell ； cancer treatment vaccine

參考文獻

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