Forced degradation of cell-based medicinal products guided by flow imaging microscopy
| Autor: | Adam D. Grabarek, Wim Jiskoot, T. Menzen, Andrea Hawe, K. Pike-Overzet |
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| Jazyk: | angličtina |
| Rok vydání: | 2021 |
| Předmět: |
Microscopy
Cell Survival Chemistry Dimethyl sulfoxide Cell Cell- and Tissue-Based Therapy Temperature Pharmaceutical Science General Medicine Jurkat cells Flow imaging Machine Learning Jurkat Cells chemistry.chemical_compound medicine.anatomical_structure Forced degradation medicine Biophysics Humans Dimethyl Sulfoxide Viability assay Fetal bovine serum Biotechnology |
| Zdroj: | European Journal of Pharmaceutics and Biopharmaceutics, 167, 38-47 European Journal of Pharmaceutics and Biopharmaceutics |
| Popis: | Cell-based medicinal products (CBMPs) offer ground-breaking opportunities to treat diseases with limited or no therapeutic options. However, the intrinsic complexity of CBMPs results in great challenges with respect to analytical characterization and stability assessment. In our study, we submitted Jurkat cell suspensions to forced degradation studies mimicking conditions to which CBMPs might be exposed from procurement of cells to administration of the product. Flow imaging microscopy assisted by machine learning was applied for determination of cell viability and concentration, and quantification of debris particles. Additionally, orthogonal cell characterization techniques were used. Thawing of cells at 5 °C was detrimental to cell viability and resulted in high numbers of debris particles, in contrast to thawing at 37 °C or 20 °C which resulted in better stability. After freezing of cell suspensions at -18 °C in presence of dimethyl sulfoxide (DMSO), a DMSO concentration of 2.5% (v/v) showed low stabilizing properties, whereas 5% or 10% was protective. Horizontal shaking of cell suspensions did not affect cell viability, but led to a reduction in cell concentration. Fetal bovine serum (10% [v/v]) protected the cells during shaking. In conclusion, forced degradation studies with application of orthogonal analytical characterization methods allow for CBMP stability assessment and formulation screening. |
| Databáze: | OpenAIRE |
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