Characterising a PDMS based 3D cell culturing microfluidic platform for screening chemotherapeutic drug cytotoxic activity

Abstract

Three-dimensional (3D) spheroidal cell cultures are now recognised as better models of cancers as compared to traditional cell cultures. However, established 3D cell culturing protocols and techniques are time-consuming, manually laborious and often expensive due to the excessive consumption of reagents. Microfluidics allows for traditional laboratory-based biological experiments to be scaled down into miniature custom fabricated devices, where cost-effective experiments can be performed through the manipulation and flow of small volumes of fluid. In this study, we characterise a 3D cell culturing microfluidic device fabricated from a 3D printed master. HT29 cells were seeded into the device and 3D spheroids were generated and cultured through the perfusion of cell media. Spheroids were treated with 5-Fluorouracil for five days through continuous perfusion and cell viability was analysed on-chip at different time points using fluorescence microscopy and Lactate dehydrogenase (LDH) assay on the supernatant. Increasing cell death was observed in the HT29 spheroids over the five-day period. The 3D cell culturing microfluidic device described in this study, permits on-chip anti-cancer treatment and viability analysis, and forms the basis of an effective platform for the high-throughput screening of anti-cancer drugs in 3D tumour spheroids.

Spheroid-on-chip microfluidic technology for the evaluation of the impact of continuous flow on metastatic potential in cancer models in vitro

“…Instead, spheroids could now be loaded directly into the spheroid well. This also accommodated the use of larger sized spheroids; spheroids generated on-chip for high-throughput applications are generally very small, 24,40 and we wished to continue using large spheroids to recapitulate the relevant in vivo zoning of tumors. Therefore, spheroids were generated off-chip with seeding at higher densities and transferred to the device after 96 h. To maintain a closed, sterile environment in the spheroid microwell, PDMS plugs were utilized to close the entry port hole after a spheroid had been loaded onto the chip [ Fig.…”

3D-Printed Collagen Scaffolds Promote Maintenance of Cryopreserved Patients-Derived Melanoma Explants

“…Monolayer culture is not sufficient to manifest all of the many complex properties of the melanoma microenvironment in vivo [ 20 , 21 , 22 ]. In vitro 3D organotypic melanoma spheroids are instead better able to portray the in vivo architecture of malignant melanoma [ 23 , 24 ]. However, nutrient and/or oxygen consumption and metabolite gradients in the outer and inner halves of the viable rim in in vitro 3D organotypic melanoma spheroids become insufficient to maintain the growth over a long culture.…”

Microvascular fragment spheroids: Three-dimensional vascularization units for tissue engineering and regeneration

“…Moreover, they may also be suitable to study the pro- and anti-angiogenic effects of novel drugs by means of the spheroids-on-a-chip technology. 41 In this study, we used the liquid overlay technique for the fabrication of spheroids, which allows a precise control of the spheroid size by using defined numbers of SVF and MVF. However, this technique is time-consuming and, thus, difficult to implement into clinical practice.…”