Multiple Analyses of Tumor Invasion Based on an Integrated Microfluidic System with Bi-directional Solute Concentration Gradient †

doi:10.7503/cjcu20190206

Abstract

Abstract:

A simple integrated microfluidic system with continuous 2D concentration gradient for multiple evaluation of tumor invasion was developed. The continuous bi-directional gradient was established vertically by solute diffusion and horizontally by Christmas tree-shaped network. The microfluidic system managed to quantitatively evaluate different invasive potentials of different cell lines(HEK-293, MCF-7, SGC-7901) and effects on tumor invasion by multiple factors. Human gastric cancer SGC-7901 cells were driven to invade into matrix under vertical FBS(Fetal bovine serum) gradient and revealed an increased invasion distance with the obvious concentration gradient. Given the continuous bi-directional gradient of FBS and Staurosporine, the invasion distance and increased number of human gastric cancer SGC-7901 cells were gradually inhibited along with the increase of drug concentration. In conclusion, the established bi-directional gradient microfluidic system provides multifunctional application in investigation of tumor invasion, cell interaction under complex environmental factors or combined chemotherapy.

Key words: Microfluidic, Concentration gradient, Tumor, Invasion, Drug screening

CLC Number:

O657

TrendMD:

Jie WANG,Yu ZHANG,Min YU,Jin FANG. Multiple Analyses of Tumor Invasion Based on an Integrated Microfluidic System with Bi-directional Solute Concentration Gradient ^†[J]. Chem. J. Chinese Universities, 2019, 40(12): 2494.

Figures/Tables 8

Fig.1 Bi-directional gradient microfluidic system (A) The schematic illustration of microfluidic device; (B) generation principle of the bi-directional gradient device.

Fig.2 Formation and maintenance of concentration gradients at different flow rates Red, blue water-soluble dye and clear ultrapure water were pumped into device simultaneously. Concentration gradient images were taken under stereomicroscope at a series of time points.

Fig.3 Horizontal and vertical concentration gradient at the flow rate of 3 μL/h Gray value of horizontal red(A), horizontal blue(B) and vertical(C) concentration gradient at 24 h was converted by Image J. 1. Cell culture channel; 2. matrix channel; 3. conditional medium channel.

Fig.4 Quantitative evaluation of horizontal and vertical concentration gradient at the flow rate of 3 μL/h (A) Horizontal red; (B) horizontal blue; (C) vertical.

Fig.5 Invasiveness detection of different cell lines Three types of cell lines(HEK-293, MCF-7, SGC-7901) were cultured in the Matrigel-loaded device under a 0—10% FBS gradient.

Fig.6 Invasiveness evaluation of different cell lines Invasiveness of different cell lines(HEK-293, MCF-7, SGC-7901) was assessed quantitatively according to the invasion distance of leading cells.

Fig.7 Invasion profile of tumor cells under bi-directional gradient (A) Human gastric cancer SGC-7901 cells were cultured and driven to invade into matrix by bi-directional FBS gradient. The invasion was imaged at sequential sites. (B) Under horizontal Staurosporine gradient and vertical FBS gradient, cultured SGC-7901 cells were driven to invade into matrix. The invasion profile was sequential imaged.

Fig.8 Quantitative invasion analysis of tumor cells under bi-directional gradient (A) Quantitative analysis of invasion distances at different sites over a 7 d period of time. The corresponding drug concentrations at different sites were also demonstrated. (B) Invasion distances at different sites were quantified on the third day and the fifth day. (C) Cell numbers within channels were also quantified accordingly for proliferation evaluation. (A) Drug concentration gradient/(nmol·L-1): a. 14; b. 16; c. 25; d. 45; e. 50; f. 60; g. 80; h. 90; i. 95; j. 110; k. 125; l. 135; m. 140; n. 160; o. 170; p. 200.

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