The Application Of Microfluidic Chip On The Drug Sensitivity Test Of Lung Cancer With3D Co-culture Mold

Objectives: Lung cancer is the leading cause of cancer death worldwide.Chemotherapy is still a major strategy for patients of lung cancer now, however theeffective rate of chemotherapy is only around20-40%. It always fails in chemotherapyand individualized treatment in clinic because of lack of accurate and rapid drugsensitivity test. The existing drug sensitive test in vitro is insufficient with2D, a single,static cell culture mode, and it is far away from the real condition of solid tumors invivo. Microfluidic chip (lab-on-a-chip), with its miniaturization, integration,high-throughput, short reaction time, less reagent consumption and other advantages isrecognized as one of the main technology platform of system biology research. Themicro channel size and multidimensional network formed a relative closed cell spacewhich is similar with characteristics, especially suitable for cell culture, and cancontinue to supply nutrients cells, to simulate the body tumor microenvironment. In thiswork, we build a new drug sensitivity test of lung cancer cells with the mold of3Dco-culture using an integrated microfluidic device. The purpose of this work lies inusing the microfluidic chip to fabricate a new drug sensitivity platform for3Dco-culturing cells and hopefully to find a higher accuracy, a stronger clinical reliabilityand more guiding for chemotherapy drug sensitivity in vitro and to investigate theapplication of microfluidic chip in individualized treatment of lung cancer.Methods: In this study, the microfluidic chip system which includes amicrofluidic chip and two MS26injection pumps was built for drug sensitivity analysisof lung cancer cell lines and fresh tissues. The main components of the microchip werecell culture chambers and concentration gradient generator (CGG). Cells and BMEmixture was infused into the3D cell culture chambers via the inlet with the connectionof a pump. Various anticancer drugs were pumped into the chambers at different concentrations by CGG after cells co-culture. Cell staining and cell apoptosis assaywere followed by a confocal laser scanning microscope after the treatments withdifferent drugs. Firstly, the human non-small cell lung cancer cell line SPCA-1wasassayed on the microfluidic chip for drug sensitivity and the traditional method MTTassay was also needed in parallel. Then SPCA-1and the human fibroblast lung cells(HFL-1) were co-cultured to investigate the effects of interstitial cells on drugsensitivity of lung cancer cells. Finally, the new platform was applied to the individualtreatment of lung cancer. The primary lung cancer cells from fresh tissue of a lungcancer patient were also cultured with3D mode in the microfluidic chip for drugsensitivity. Subsequently, screening the best anticancer drugs for individuals andproviding a basis for clinical individual therapy.Results: In this study, we have designed and built a microfluidic chip on the drugsensitivity test of lung cancer cells with the mold of3D co-culture successfully. Themicrofluidic chip concentration gradient generator can produce a5:6:8drugconcentration. In cell culture unit we have completed a single cell, different cells andlung cancer tissues with the mold of3D co-culture. The results of drug sensitivitydetected by cell apoptosis staining on microchip were consistent with the traditionalMTT assay. For interstitial cells, we found that the apoptosis rate of lung cancer cellsdecreased under the action of the interstitial cells compared to lung cancer cell itself, itindicated that the interstitial cells existed around lung cancer cells could decrease thedrug sensitivity. Chemotherapy drug screening was evaluated with primary lung cancercells from a surgical specimen of a lung cancer patient by the use of the microfluidicchip3D co-culture drug sensitivity platform.Conclusion: This study has successfully fabricated a microfluidic3D co-culturedrug sensitivity platform,which was available for lung cancer cell lines and primarylung cancer cells from tissues3D cultures had provide a fast, efficient and accurate newdrug testing platform for clinical individualized treatment.