Temperature-Dependent Cell Detachment on Pluronic Gels

Abstract

Cell culture on gels made of poly(ethylene oxide) and poly(propylene oxide) (Pluronic), which has a lower critical solution temperature around 30 degrees C, could be performed for 48 h. However, the Pluronic gels were highly hydrophilic and tended to dissolve in the culture medium. We achieved temperature-dependent detachment of cells from Pluronic gels containing or lacking extracellular matrix (ECM) by cooling the gels to 4 degrees C. Using normal human umbilical vein endothelial cells (HUVECs) grown on and released from Pluronic gels lacking ECM, we further found that the expression ratio of the surface markers CD34 and CD105 was twofold higher than for cells grown on polystyrene and removed with trypsin. In addition, the expression ratios for CD34 and CD105 on HUVECs cultivated on the Pluronic gels containing higher concentrations of ECM were lower, which may be due to ECM coating of the cell surface and, thus, interference with antibody binding. In summary, temperature-dependent detachment of cells from Pluronic gels allows the isolation of cells under mild conditions. This can be a powerful tool for surface marker analysis by flow cytometry.

Reverse Phase Compatible TLC‐Bioautography for Detection of Tyrosinase Inhibitors

“…They can form temperature dependent micelles that facilitate solubilisation of hydrophobic compounds. Water solutions of poloxamer show thermo-reversible gelation behaviour (Bohorquez et al, 1999;Higuchi et al, 2005): they are sols below room temperature and become gels at 37°C (Ruel-Gariépy and Leroux, 2004). In addition, poloxamer gels can help to preserve enzymes from alkaline stress and aggregation, and to increase their thermal stability (Pucciarelli et al, 2012).…”

A reversed‐phase compatible thin‐layer chromatography autography for the detection of acetylcholinesterase inhibitors

“…Micelles formation allows solubilization of hydrophobic compounds, making these copolymers attractive for delivering poorly water‐soluble compounds. Water solutions of Poloxamer 407 with concentrations of 20% w/v or above, show interesting thermoreversible gelation behavior . They are sols below room temperature and get converted to gels at body temperature (37.2°C) .…”

Engineered extracellular matrices with cleavable crosslinkers for cell expansion and easy cell recovery

“…Thermoresponsive polymers [31] have been used to recover cell sheets from an elastin polymer-coated inert membrane [46], from cells cultured on poly( N -isopropylacrylamide) (PIPAAm) grafted PEG surfaces [47, 48], or from cells cultured on pluronic-coated surfaces [49]. These novel surfaces allow the cultivation of cells without using enzymes by utilizing the thermoresponsive phase transition property of PNIPAAm [50].…”