Abstract
ABSTRACTHigh quality cell cultures require reliable laboratory practices. Today's small-scale in vitro cell culture format is dominated by circular topology vessels, with the inherent disadvantage of secondary flow induced each time the cell cultures are repositioned. The secondary flow generates uneven sedimentation and adherence that negatively impacts cell culture quality. Here we show a modification of the circular culture vessel that abrogates these disturbances. Cell culture wells were augmented with a central column to diminish secondary flow. Human carcinoma cell lines (BeWo, JEG-3), mesenchymal stem cells [human umbilical cord perivascular cells (HUCPVC)] and mouse embryonic fibroblasts (MEF) were cultured in both column-augmented and regular culture wells. Human carcinoma cell cultures showed even cell densities and significantly more viable cells in column-augmented vessels. In FTM HUCPVC cultures, cell surface MSC marker (CD90, CD105) expression and cell differentiation-related gene expression patterns were significantly more homogeneous in column-augmented vessels. MEF cells in column-augmented culture vessels showed a more consistent expression of IGF-1. Column-augmented cell culture vessels significantly improve the homogeneity of adherent cell cultures by mitigating the adverse effect of the secondary flow.This article has an associated First Person interview with the first author of the paper.
Highlights
Principles of small-scale adherent cell cultures Small-scale cell culture experiments most commonly aim to characterize the identity and potential of adherent eukaryotic cells
The cylindrical culture vessel is still commonly used in cell biology laboratories worldwide
The work presented here demonstrates that fluid mechanics, in particular the induction of secondary flow, in the small-scale, circular culture vessels can significantly impact a cell culture’s quality
Summary
Principles of small-scale adherent cell cultures Small-scale cell culture experiments most commonly aim to characterize the identity and potential of adherent eukaryotic cells. The principles of small-scale cultures date back to Julius Richard Petri’s research in the 1880s (Petri, 1887). Petri’s microbiological practices introduced the cylindrical cell culture vessel that has been in use in various. Eukaryotic cell research adopted this cell culture vessel topology and various sizes of the cylindrical cell culture dishes are utilized worldwide
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