Evaluation of cell viability and metabolic activity of a 3D cultured human epidermal model using a dynamic autoradiographic technique with a PET radiopharmaceutical

Abstract

Quality control of tissues and organs for transplant is important to confirm their safety and effectiveness for regenerative medicine. However, quality evaluation is only carried out using a limited range of inspection criteria, because many of the available evaluation tests are invasive. In order to explore the potential of 2-[ 18 F]fluoro-2-deoxy-D-glucose ([ 18 F]FDG)-bioradiography as a non-invasive test for estimation of the safety, soundness, and effectiveness of tissues for transplantation, [ 18 F]FDG uptake and cell viability or metabolism were investigated using a reconstructed human epidermal model (RHEM). We developed an imaging system, and suitable bioradiographic image acquisition conditions and its effectiveness were investigated. [ 18 F]FDG uptake increased in agreement with DNA content as a marker of cell numbers and for histological assessment during cell proliferation and keratinization. [ 18 F]FDG uptake was significantly decreased in good agreement with the viability of tissues used with various hazardous chemical treatments. [ 18 F]FDG uptake by the tissues was decreased by hypothermia treatment and increased by hypoxia treatment while maintaining cell viability in the tissue. Therefore, [ 18 F]FDG-bioradiography can be useful to estimate cell viability or metabolism in this RHEM. This method might be utilized as a non-invasive test for quality evaluation of tissues for transplantation.

Cyclodextrin Diethyldithiocarbamate Copper II Inclusion Complexes: A Promising Chemotherapeutic Delivery System against Chemoresistant Triple Negative Breast Cancer Cell Lines

“…The cytotoxic effect of SBE20 and HP20 freeze-dried formulations on breast cancer cells was assessed in vitro. Living cells, indicated by metabolic activity, were estimated using an MTT assay and presented as a percentage of the non-treated cells [ 19 ]. MDA-MB-231 and MDA-MB-231 PAC10 cells were seeded in 96-well plates at a density of 10 × 10 3 cells for MDA-MB-231 and 5 × 10 3 cells for MDA-MB-231 PAC10 in Dulbecco’s modified Eagle’s medium (DMEM) with 10% FBS, 1 mM sodium pyruvate, 2 mM L-glutamine, 50 IU/mL penicillin, 50 μg/mL streptomycin and 0.1 mM non-essential amino acids.…”

Organ Cultures for Retinal Diseases

“…There are numerous bioassays to investigate cell metabolism, cell death, or proliferation. Many of these assays can also be performed on retinal tissue, such as the measurement of reactive oxygen species (ROS) production from hypoxia-stressed retinal explants (Maliha et al, 2019;Sasaki et al, 2019; Figure 1).…”

Advancements in Assessments of Bio-Tissue Engineering and Viable Cell Delivery Matrices Using Bile Acid-Based Pharmacological Biotechnologies

“…A variety of chemical reagents can be used to culture islet/cell containing microcapsules for short periods of time in order to gently rupture the islets/cells [ 82 , 83 , 84 ]. The common method is to place a set number of microcapsules (routinely around 50–100) into a sterile container containing 1 mL PBS or culture media, be it RPMI, CMRL or DMEM, supplemented with 0.25% trypsin and 0.5 mM EDTA and incubating at 37 °C CO 2 (95% humidified air and 5% CO 2 ) with gentle shaking periodically [ 74 ].…”

“…The change in colour (if present) can be measured photometrically [ 85 , 86 , 87 ]. Despite its ability to determine viability, the MTT assay does have drawbacks, including that microcapsules put through the assay are no longer viable, and the that increased incubation with MTT reagents causes improved sensitivity in results, but is more cytotoxic [ 84 ]. Despite known limitations, the MTT assay is still commonly used in the field of islet cell microencapsulation [ 5 ].…”