Enhanced production of carcinoembryonic antigen by CW-2 cells cultured on polymeric membranes immobilized with extracellular matrix proteins

Hara, Mariko; Adachi, Shin; Higuchi, Akon

doi:10.1163/156856203321142588

Cited by 6 publications

(8 citation statements)

References 25 publications

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“…The cell number was characterized by observation of the cells using an inverted microscope (Diaphoto TMD300, Nikon Co., Tokyo) equipped with a charge-coupled device (CCD) video camera, ARGUS 20 (Hamamatsu Photonics K.K.) and a temperature-regulated box. − …”

Section: Methodsmentioning

confidence: 99%

Temperature-Dependent Cell Detachment on Pluronic Gels

et al. 2005

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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.

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Section: Methodsmentioning

confidence: 99%

Temperature-Dependent Cell Detachment on Pluronic Gels

et al. 2005

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“…Then, the numbers of residual cells were counted using an inverted microscope (Diaphoto TMD300, Nikon) equipped with a CCD video camera, ARGUS 20 (Hamamatsu Photonics K.K. ), digital camera (Camedea, Olympus), and a temperature‐regulated box 19–22. The cell number was obtained from the averages of our different experiments on each well using four independent wells prepared from the same conditions (totally n = 16).…”

Section: Methodsmentioning

confidence: 99%

Temperature‐induced cell detachment on immobilized pluronic surface

Higuchi

Aoki

Yamamoto

et al. 2006

J Biomedical Materials Res

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The Pluronic F68 and F127, a triblock copolymer of ethylene oxide and propylene oxide, was activated using carbonyldiimidazole (CDI), and CDI-activated Pluronic F68 and F127 was subsequently immobilized on the surface of a poly-L-lysine-coated polystyrene tissue culture flask. Cell culture was performed on the Pluronic-immobilized flask. The morphology of fibroblasts (L929 cells) on the Pluronic F127-immobilized flask was mainly spherical, and showed less spreading behavior than that on the Pluronic F68-immobilized flask and conventional tissue culture flask. This observation indicates that L929 cells on Pluronic F127-immobilized flasks were cultured in a bio-inert environment. L929 cells were successively detached from both Pluronic F127-immobilized flask and Pluronic F68-immobilized flask by cooling the flask to 4-15 degrees C. This detachment is due to the hydration and dehydration properties of Pluronic, depending on the temperature. Umbilical cord blood was cultured in the Pluronic F127-immobilized and conventional polystyrene tissue culture flasks at 37 degrees C. The expression ratio of surface markers on hematopoietic stem cells (CD34 and CD133) cultured in the Pluronic F127-immobilized flask was significantly higher than that of the cells in polystyrene tissue culture flask.

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“…Cell Cultivation of L929 Cells. L929 cells (Dainippon Seiyaku Co., Ltd.), derived from a mouse connective tissue fibroblast, were maintained in RPMI 1640 media (JRH Bioscience, Lenexa, KS) supplemented with 25 mg/L streptomycin sulfate, 3.5 mg/L benzylpenicillin potassium, and 10% fetal bovine serum (FBS, JRH Bioscience, Lenexa, KS) in 5% CO 2 atmosphere at 37 °C . The cell lines were used between passage numbers 10 to 20 in the following experiments.…”

Section: Methodsmentioning

confidence: 99%

“…L929 cells in suspension (cell densities 5 × 10 4 /cm 2 and 2 mL) were injected into the Pluronic-immobilized flasks and tissue culture flasks, and they were incubated in the CO 2 incubator in 5% CO 2 atmosphere at 37 °C. The numbers of cells were counted using an inverted microscope (Diaphoto TMD300, Nikon Co.) equipped with a CCD video camera, ARGUS 20 (Hamamatsu Photonics K. K.), digital camera (Camedea, Olympus Co.), and a temperature-regulated box. − The cell number was obtained from the average of four different experiments on each well using four independent wells prepared under the same conditions (total n = 16). The dynamic morphology of the cells was also recorded using the inverted microscope equipped with the CCD video camera and temperature-regulated box.…”

Section: Methodsmentioning

confidence: 99%

“…L929 cells (Dainippon Seiyaku Co., Ltd.), derived from a mouse connective tissue fibroblast, were maintained in RPMI 1640 media (JRH Bioscience, Lenexa, KS) supplemented with 25 mg/L streptomycin sulfate, 3.5 mg/L benzylpenicillin potassium, and 10% fetal bovine serum (FBS, JRH Bioscience, Lenexa, KS) in 5% CO 2 atmosphere at 37 °C. 20 The cell lines were used between passage numbers 10 to 20 in the following experiments. L929 cells in suspension (cell densities 5 × 10 4 /cm 2 and 2 mL) were injected into the Pluronic-immobilized flasks and tissue culture flasks, and they were incubated in the CO2 incubator in 5% CO2 atmosphere at 37 °C.…”

Section: Methodsmentioning

confidence: 99%

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Bioinert Surface of Pluronic-Immobilized Flask for Preservation of Hematopoietic Stem Cells

et al. 2006

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The bioinert materials on which cells do not proliferate, differentiate, nor de-differentiate should be useful for the culture and preservation of stem cells. The Pluronic F127, a triblock copolymer of ethylene oxide, and propylene oxide was activated using carbonyldiimidazole (CDI), and CDI-activated Pluronic was subsequently immobilized on the surface of a lysine-coated polystyrene tissue culture flask. The morphology of fibroblasts (L929 cells) on the Pluronic-immobilized flask was spherical, and did not show spreading behavior. This observation indicates that L929 cells on the Pluronic-immobilized flask were cultured in a bioinert environment. The expression ratio of surface markers on hematopoietic stem cells (CD34 and CD133) cultured in the Pluronic-immobilized flask was significantly higher than that in polystyrene tissue culture flask and commercially available bioinert flask (i.e., low cell binding cultureware). This is caused by the existence of hydrophilic segments of Pluronic F127 on the Pluronic-immobilized flask.

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Enhanced production of carcinoembryonic antigen by CW-2 cells cultured on polymeric membranes immobilized with extracellular matrix proteins

Cited by 6 publications

References 25 publications

Temperature-Dependent Cell Detachment on Pluronic Gels

Temperature-Dependent Cell Detachment on Pluronic Gels

Temperature‐induced cell detachment on immobilized pluronic surface

Bioinert Surface of Pluronic-Immobilized Flask for Preservation of Hematopoietic Stem Cells

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