The importance of cell culture parameter standardization: an assessment of the robustness of the 2102Ep reference cell line
| Autor: | James Willard Tonderai Kusena, Jenna Rebekah James, Samantha Loiuse Wilson, Robert Thomas, Adam James Studd, Maryam Shariatzadeh |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2021 |
| Předmět: |
0301 basic medicine
Biomedical Research Standardization Computer science reference cell lines Cytological Techniques Cell- and Tissue-Based Therapy cell growth rates Bioengineering Applied Microbiology and Biotechnology Cell Line 03 medical and health sciences 0302 clinical medicine Humans quality control metabolites Cell Proliferation Robustness (evolution) Reference cell General Medicine Reference Standards Reliability engineering 030104 developmental biology defined cell therapy manufacturing 030217 neurology & neurosurgery Biomarkers TP248.13-248.65 Research Article Research Paper Biotechnology |
| Zdroj: | Bioengineered, Vol 12, Iss 1, Pp 341-357 (2021) Bioengineered article-version (VoR) Version of Record |
| ISSN: | 2165-5987 2165-5979 |
| Popis: | Work undertaken using the embryonic carcinoma 2102Ep line, highlighted the requirement for robust, well-characterized and standardized protocols. A systematic approach utilizing ‘quick hit’ experiments demonstrated variability introduced into culture systems resulting from slight changes to culture conditions (route A). This formed the basis for longitudinal experiments investigating long-term effects of culture parameters including seeding density and feeding regime (route B).Results demonstrated that specific growth rates (SGR) of passage 59 (P59) cells seeded at 20,000 cells/cm2 and subjected to medium exchange after 48h prior to reseeding at 72h (route B2) on average was marginally higher than, P55 cells cultured under equivalent conditions (route A1); whereby SGR values were (0.021±0.004) and (0.019±0.004). Viability was higher in route B2 over 10 passages with average viability reported as (86.3%±8.1) compared to route A1 (83.3±8.8). The metabolite data demonstrated both culture route B1 (P57 cells seeded at 66,667 cells/cm2) and B2 had consistent-specific metabolite rates (SMR) for glucose, but SMR values of route B1 was consistently lower than route B2 (0.00001 mmol, cell-1.d-1 and 0.000025).Results revealed interactions between phenotype, SMR and feeding regime that may not be accurately reflected by growth rate or observed morphology. This implies that current schemes of protocol control do not adequately account for variability, since key cell characteristics, including phenotype and SMR, change regardless of standardized seeding densities. This highlights the need to control culture parameters through defined protocols, for processes that involve culture for therapeutic use, biologics production, and reference lines. GRAPHICAL ABSTARCT |
| Databáze: | OpenAIRE |
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