Bags versus flasks: a comparison of cell culture systems for the production of dendritic cell–based immunotherapies

Abstract

In recent years, cell-based therapies targeting the immune system have emerged as promising strategies for cancer treatment. This review summarizes manufacturing challenges related to production of antigen presenting cells as a patient-tailored cancer therapy. Understanding cell-material interactions is essential because in vitro cell culture manipulations to obtain mature antigen-producing cells can significantly alter their in vivo performance. Traditional antigen-producing cell culture protocols often rely on cell adhesion to surface-treated hydrophilic polystyrene flasks. More recent commercial and investigational cancer immunotherapy products were manufactured using suspension cell culture in closed hydrophobic fluoropolymer bags. The shift to closed cell culture systems can decrease risks of contamination by individual operators, as well as facilitate scale-up and automation. Selecting closed cell culture bags over traditional open culture systems entails different handling procedures and processing controls, which can affect product quality. Changes in culture vessels also entail changes in vessel materials and geometry, which may alter the cell microenvironment and resulting cell fate decisions. Strategically designed culture systems will pave the way for the generation of more sophisticated and highly potent cell-based cancer vaccines. As an increasing number of cell-based therapies enter the clinic, the selection of appropriate cell culture vessels and materials becomes a critical consideration that can impact the therapeutic efficacy of the product, and hence clinical outcomes and patient quality of life.

Plant-Based Scaffolds Modify Cellular Response to Drug and Radiation Exposure Compared to Standard Cell Culture Models

“…Therefore, we also assessed the stiffness of different decellularized plants, including spinach, tomato, aquatic plant, basil, A. Borealis leaves and lucky bamboo (Figure 6). Our results showed that resulting scaffolds displayed a broad range of stiffness, with Young's modulus from 1.7 ± 0.3 (A. Borealis) to 1767 ± 1260 kPa (lucky bamboo), thus matching stiffness of main human tissues (Cox and Erler, 2011;Skardal et al, 2013;Handorf et al, 2015;Yang et al, 2017;Fekete et al, 2018) and confirming the potential use of cellulose scaffolds produced from plants decellularization to provide appropriate matrixes for reproducing the unique stiffness of specific tissue, health condition or disease progression. For example, fibrotic tissue or tumor are known to be stiffer than normal tissue (Cross et al, 2007;Liu et al, 2015), and, according to their stiffness, leaf scaffolds could be selected to mimic appropriate disease or normal microenvironment.…”

Closing the system: production of viral antigen-presenting dendritic cells eliciting specific CD8+ T cell activation in fluorinated ethylene propylene cell culture bags

“…Further, Mo-DC have recently successfully been used to induce viral-specific T cell expansion in virus-naïve donors [36]. This technology could be valuable for a variety of applications such as cancer immunotherapy [2,13], the expansion of tumor or viral reactive T cells, to treat, respectively, cancer or infection-prone immunocompromised patients [37,38] as well as expanding other tolerogenic cells such as regulatory T cells (Tregs) in the context of auto-immunity or graft rejection [39].…”

“…As recently reviewed by Fekete et al . [ 13 ], the transition from functionally-open TCPS plates to closed systems such as FEP or polyolefin bags leads to a concurrent transition in material properties including gas permeability, mechanical properties, surface topography, surface chemistry and surface wettability. This may affect protein adsorption profiles and resulting changes in the cell microenvironment which may impact Mo-DC cell fate decisions, as observed with other therapeutic cells [ 14 ].…”

“…Nevertheless, most translational research studies are performed in tissue culture polystyrene (TCPS)-based plates or flasks of much smaller volumes (typically ≤ 10 mL) and conducted in rigid tissue culture polystyrene (TCPS)-based plates or flasks. As recently reviewed by Fekete et al [13], the transition from functionally-open TCPS plates to closed systems such as FEP or polyolefin bags leads to a concurrent transition in material properties including gas permeability, mechanical properties, surface topography, surface chemistry and surface wettability. This may affect protein adsorption profiles and resulting changes in the cell microenvironment which may impact Mo-DC cell fate decisions, as observed with other therapeutic cells [14].…”

Hurdles to uptake of mesenchymal stem cells and their progenitors in therapeutic products

“…a culture flask requiring opening for media exchange within a flow cabinet) or closed (e.g. bag culture with automated media exchange on benchtop) [140,141].…”