A solution to prevent secondary flow in adherent cell cultures

Abstract

High 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. .

Development and Characterization of Alkaline Phosphatase-Positive Human Umbilical Cord Perivascular Cells

“…Greater quantities of immature MSCs can be obtained from fetal appendages such as Wharton’s jelly in the umbilical cord and placental amniotic membrane [ 10 , 11 , 12 ]. Human umbilical cord perivascular cells (HUCPVCs) harvested from human umbilical cord perivascular tissue are an MSC population and have been used as a cell source for dermal wound healing [ 13 , 14 ] and other MSC studies [ 15 , 16 , 17 ]. HUCPVCs area mesenchymal cell population with a high colony-forming unit-fibroblast frequency at harvest is 1:300, which is considerably higher than that of bone marrow, at 1:100,000.…”

First person – Peter Szaraz

“…ABSTRACTFirst Person is a series of interviews with the first authors of a selection of papers published in Biology Open, helping early-career researchers promote themselves alongside their papers. Peter Szaraz is first author on ‘A solution to prevent secondary flow in adherent cell cultures’, published in BiO. Peter is a Research Associate in the lab of Clifford L. Librach at the University of Toronto, Canada, investigating in vitro techniques to test and preserve the regenerative potential of cell therapy candidates.…”