Validation of a novel perfusion bioreactor system in cancer research

Abstract

Development of drugs is a complex, time- and cost-consuming process due to the lack of standardized and reliable characterization techniques and models. Traditionally, drug screening is based on in vitro analysis using two-dimensional (2D) cell cultures followed by in vivo animal testing. Unfortunately, application of the obtained results to humans in about 90 % of cases fails. Therefore, it is important to develop and improve cell-based systems that can mimic the in vivo-like conditions to provide more reliable results. In this paper, we present development and validation of a novel, user-friendly perfusion bioreactor system for single use aimed for cancer research, drug screening, anti-cancer drug response studies, biomaterial characterization, and tissue engineering. Simple design of the perfusion bioreactor provides direct medium flow at physiological velocities (100?250 ?m s-1) through samples of different sizes and shapes. Biocompatibility of the bioreactor was confirmed in short term cultivation studies of cervical carcinoma SiHa cells immobilized in alginate microfibers under continuous medium flow. The results have shown preserved cell viability indicating that the perfusion bioreactor in conjunction with alginate hydrogels as cell carriers could be potentially used as a tool for controlled anti-cancer drug screening in a 3D environment. [Projects of the Serbian Ministry of Education, Science and Technological Development, Grant no. 451-03-68/2020-14/200135 and Grant no. 451-03-68/2020-14/ 200042]

Development and Validation of a Long-Term 3D Glioblastoma Cell Culture in Alginate Microfibers as a Novel Bio-Mimicking Model System for Preclinical Drug Testing

“…The extrusion process had several optimization steps in order to obtain sustainable, long-term 3D cell culture. Based on previously published data, we decided to mix 2% w/v sodium alginate solution with the cell suspension to obtain a final concentration of 1.5% w/v alginate [20]. Further, the transport of oxygen and nutrients by diffusion through alginate hydrogels is limited to 200 µm [21].…”

“…Another important factor in the optimization study is the initial number of cells used to obtain 3D cultures. Previous studies with alginate scaffolds reported initial inoculation numbers from 1 × 10 5 to 1 × 10 7 cells, depending on the experimental purposes [20,[23][24][25][26]. In our model system, we used three different inoculation densities of U87 cells (1 × 10 6 , 2 × 10 6, and 4 × 10 6 cells/mL alginate), and all of them gave rise to the 3D culture that preserved proliferative potential during 28 days.…”

Chemical engineering methods in analyses of 3D cancer cell cultures: Hydrodinamic and mass transport considerations

“…The single-use perfusion bioreactor system "3D Perfuse" (Innovation Center of the Faculty of Technology and Metallurgy, Belgrade, Serbia) was used for cultivation of alginate microfibers with immobilized rat glioma C6 cells. As before, the system consisted of a chamber, two 3-way stopcocks, a medium reservoir and a silicone tubing loop serving for gas exchange [41]. In the present experimental setup 0.5 g of wet microfibers with immobilized rat glioma C6 cells, cultured statically for 8 days, was placed in the bioreactor chamber (0.8 cm inner diameter, 4 cm height) formed by a piece of silicone tubing and the system was filled with 15 cm 3 of the culture medium.…”

Interdisciplinary crossover for rapid advancements - collaboration between medical and engineering scientists with the focus on Serbia

“…Studies of novel macroporous, composite scaffolds based on gellan gum and bioactive glass particles have shown that the fluid flow in perfusion bioreactors significantly promoted formation of a mineral phase as compared to static conditions, which could be related to scaffold implantation in vascularized tissues [24]. Finally, an integrative approach to optimization of biomaterial properties and operating conditions in biomimetic bioreactors may provide tools for evaluation of new drugs and treatment procedures on 3D models of human tissues, as well as for development of personalized medical therapies, and thus contribute to decrease the level of needed animal experimentation [29].…”