A microfluidic localized, multiple cell culture array using vacuum actuated cell seeding: integrated anticancer drug testing

Abstract

In this study, we introduced a novel and convenient approach to culture multiple cells in localized arrays of microfluidic chambers using one-step vacuum actuation. In one device, we integrated 8 individually addressable regions of culture chambers, each only requiring one simple vacuum operation to seed cells lines. Four cell lines were seeded in designated regions in one device via sequential injection with high purity (99.9%-100%) and cultured for long-term. The on-chip simultaneous culture of HuT 78, Ramos, PC-3 and C166-GFP cells for 48 h was demonstrated with viabilities of 92%+/−2%, 94%+/−4%, 96%+/−2% and 97%+/−2%, respectively. The longest culture period for C166-GFP cells in this study was 168 h with a viability of 96%+/−10%. Cell proliferation in each individual side channel can be tracked. Mass transport between the main channel and side channels was achieved through diffusion and studied using fluorescein solution. The main advantage of this device is the capability to perform multiple cell-based assays on the same device for better comparative studies. After treating cells with staurosporine or anti-human CD95 for 16 h, the apoptotic cell percentage of HuT 78, CCRF-CEM, PC-3 and Ramos cells were 36%+/−3%, 24%+/−4%, 12%+/−2%, 18%+/−4% for staurosporine, and 63%+/−2%, 45%+/−1%, 3%+/−3%, 27%+/−12% for anti-human CD95, respectively. With the advantages of enhanced integration, ease of use and fabrication, and flexibility, this device will be suitable for long-term multiple cell monitoring and cell based assays.

Probing hypoxia-induced staurosporine resistance in prostate cancer cells with a microfluidic culture system

“…The cell culture chip, shown in Figure 1A, was based on our previous vacuum actuated design (30). The bottom fluid layer and top control layer were separated by a thin PDMS layer.…”

“…The detailed description of device fabrication and operation is described in our previous work (30). The dimensions of the channels are given in Table E1 (see Supplementary Information).…”

“…The chips were injected with the cell solution and vacuum pump was connected to the vacuum line. The vacuum pump was turned on and the cells were seeded in the side channels using vacuum actuation (30). Once all the chambers were filled with medium, the vacuum was turned off.…”

“…Once all the chambers were filled with medium, the vacuum was turned off. The medium was then flowed into the chip at the rate of 0.1mL/hr, which was optimized in our previous work (30). At the flow-rate of 0.1mL/hr, the device has been shown to produce apoptosis response in the time span comparable to the Petri dish studies…”

Microfluidics for cell-based high throughput screening platforms—A review

“…Although this approach demonstrates a simple microfluidic system for cell-based cytotoxicity assays, the throughput of this system is not sufficiently high for drug discovery. Gao et al [32] used a microfluidic channel combined with vacuum actuated chambers for one-step cell seeding and anticancer drug testing.…”