Temperature-dependent cell detachment on Pluronic gels.
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.
Higuchi
Akon
A
Department of Applied Chemistry, Seikei University, 3-3-1 Kichijoji Kitamachi, Musashino, Tokyo 180-8633, Japan. higuchi@ch.seikei.ac.jp
Yamamoto
Taro
T
Sugiyama
Kaichiro
K
Hayashi
Shizue
S
Tak
Tae Moon
TM
Nakagawa
Tsutomu
T
D015703:Antigens, CD; D018952:Antigens, CD34 / Q000032:analysis; D002448:Cell Adhesion; D002469:Cell Separation / Q000379:methods*; D000071063:Endoglin; D004730:Endothelium, Vascular / Q000166:cytology; D005782:Gels; D006801:Humans; D020442:Poloxamer; D011956:Receptors, Cell Surface; D013696:Temperature; D014471:Umbilical Veins; D019010:Vascular Cell Adhesion Molecule-1 / Q000032:analysis
D015703:Antigens, CD; D018952:Antigens, CD34; C064206:ENG protein, human; D000071063:Endoglin; D005782:Gels; D011956:Receptors, Cell Surface; D019010:Vascular Cell Adhesion Molecule-1; D020442:Poloxamer