A Size-Controlled 3D Tumor Array Chip And Its Application For Antitumor Drug Testing

Cancer is a leading cause of death worldwide.In vitro testing of anticancer drugs typically involves growing cancer cell lines in monolayers on tissue culture plastic and using high-throughput testing procedures to evaluate drug efficacy.However,the monolayer culture has remained a poor predictor of whether a given drug will ultimately yield clinical benefit.The development of 3D tumor is anticipated to increase the forecast data,decrease the animal testing,and reduce the time and cost to identify new drugs.3D tumor is conventionally formed using sedimentation on non-adherent surfaces,spinner flasks,or the hanging drop method.The hanging drop method is most often relied upon due to its efficient fabrication of uniform 3D tumor and compatibility with high throughput testing instruments.Spinners offer a simple method of 3D tumor generation.However,these techniques are time-consuming,laborious,and require large amounts of reagents.Unlike the conventional cell testing methods,microfluidic platforms provide in vitro models with reduced reagent and being high-throughput.The predominance of PDMS on cell-based chips is based strongly on its gas permeability which is suitable for cell manipulation,as well as its optical clarity and biocompatibility.However,the current tumor chips only focus on formation of tumor with specific size,which lack the ability to control tumor size,as well as fail to systematically analyse tumor biology?Therefore,we designed a microfluidic device with for employing a series of sequential controls of 3D tumors,including 3D tumor formation,manipulation and on-line annlysis.We investigated optimized microfluidic device fabrication,cell localization,different sizes of HepG2/U251 tumor and its application for antitumor drug testing.we obtained following conclusions:1.The microfluidic device consisted of two layers was fabricated using soft lithography,which was applied for 3D tumor manipulation analysis.The microfluidic device was optimized fabricated with PDMS mixing ratio 20:1 and 10:1 for the two layers,which avoid the leaking of fluid.The corresponding curing time was 23 minutes and 13 minutes,respectively.2.By optimizing the cell density,flow rates and perfusion time,the precise capture and localization of HepG2 and U251 cells in the chip were realized,which laid the foundation for the further study of three-dimensional tumor culture.3.The culture of HepG2 and U251 three-dimensional tumors with different sizes in the chip was completed.The results showed that the specific size of the three-dimensional tumor prepared by with size uniformityand high tumor cell activity,which was suitable for antitumor drug analysis applications.4.Two types of antitumor drugs were analyzed based on three-dimensional tumor chip.The results showed that the cell viability decreased with the increase in drug concentration.HepG2 tumor was treated using two types of antitumor drugs of BLM and TPZ.The viability of human hepatocarcinoma cells was lower at the same concentration using TPZ drug treatments.The results show that the design of microfluidic chip can achieve different sizes of three-dimensional tumor preparation and its application for antitumor drug testing.