具人源性腫瘤異植體微型切塊之人體腫瘤晶片製程開發與驗證

Abstract

本研究為一種可使用人源腫瘤組織開發腫瘤晶片模型的微流道裝置。藉由微流體設計及操控技術在培養腔室中產生血管新生及腫瘤培養所需之培養環境，成功於體外建立具人血管的體腫瘤組織模型，將可做為藥物發展及篩選的研發平台。本研究所開發的腫瘤晶片可培養出平均血管直徑為15.6 μm的血管組織，並於培養第4天即可形成血管網絡，並於第9天即可與流道完成吻合連接。並於實驗驗證所發展出的血管組織會向腫瘤組織生長，在腫瘤尺寸範圍200-220 μm3的血管平均直徑為48.08 μm為相較於腫瘤尺寸範圍10-100 μm3的血管平均直徑30.43 μm增加1.58倍。本研究並以化療藥物進行實驗驗證，其結果顯示10倍藥物濃度的腫瘤組織壞死程度是1倍藥物濃度的1.77倍，且相較於控制組為4.6倍。最後，本研究並驗證此腫瘤模型的微環境，確認此腫瘤模型含有原人體腫瘤組織的免疫B細胞，藉此驗證腫瘤微環境的細胞多樣性，同時此腫瘤模型可將組織液取樣，以監測在培養過程中的pH值。 本研究所開發的腫瘤晶片模型提供更近似體內腫瘤環境的模型系統，將可提供具血管化的腫瘤體外模型，用以進行腫瘤藥物篩選研究，並加速藥物篩選時程及整體可靠度。

This study reports a microfluidic device that can develop tumor-on-a-chip models using human tumor tissue. Microfluidic technology is applied to create the physiological environment required for angiogenesis and tumor in the culture chamber. This microfluidic device successfully develops a human tumor model with human blood vessels. It can serve as an in vitro model system for drug development and screening. The vascular network has an average diameter of 15.6 μm, and a vascular network can be formed within 4 days. The developed vessels can form anastomosis in 9 days. It is also verified experimentally that the developed vascular tissue can grow toward the tumor tissue. The average diameter of blood vessels in the tumor size range of 200-220 μm3 is 48.08 μm, which is 1.58 times greater than the average diameter of blood vessels in the tumor size range of 10-100 μm3, which is 30.43 μm3. The capability to use this device for drug screening also is verified using a standard chemotherapy drug. The results show that the level of treated tumor tissue damage at 10X dosage is 1.77 times larger than that of 1X dosage and was 4.6 times larger than that of the control group. Finally, the microenvironment of the tumor model also is verified. It is found that the immune B cells of the original human tumor tissue can be retained in the mode, and tumor fluid can be sampled to monitor pH value during cultivation. In summary, the developed tumor-on-a-chip device provides a new model system with a more tumor-mimetic environment. It provides a reliable vascularized tumor model for speeding up tumor drug screening studies.