Immobilized culture of nonadherent cells on an oleyl poly(ethylene glycol) ether-modified surface

Kato, Koichi; Umezawa, Keitaro; Funeriu, Daniel P.; Miyake, Masato; Miyake, Jun; Nagamune, Teruyuki

doi:10.2144/03355rr01

Cited by 77 publications

(65 citation statements)

References 7 publications

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“…As a special utilization, we have already reported the application of single oleyl chain BAM-modified glass slides for immobilization of nonadherent cells (24). On the contrary, DOPE-BAM may be useful for protein anchoring in the presence of serum and is expected to be useful in vivo.…”

Section: Discussionmentioning

confidence: 99%

Rapid Protein Anchoring into the Membranes of Mammalian Cells Using Oleyl Chain and Poly(ethylene glycol) Derivatives

et al. 2004

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The cell membrane is an important interface for communication with extracellular events, and incorporation of bioactive substances, such as antibodies and receptors, into the cell membrane may enhance the potential abilities of cells. Gene manipulation, chemical modification of membrane proteins, and cell surface painting using a GPI anchor have been performed to introduce substances into cell membranes. Furthermore, many lipid anchors have also been used to modify lipid membranes such as liposomes. In this study, we have focused on developing an easy and rapid method for anchoring of substances including macromolecular proteins into the membranes of living mammalian cells. We employed a single oleyl chain derivative coupled with hydrophilic poly(ethylene glycol) (PEG90, the ethyleneoxide (EO) unit is 90) to facilitate solubilization in water. This water-soluble derivative was designated Biocompatible Anchor for Membrane (BAM). Some proteins (streptavidin, EGFP and an antibody) were coupled with BAM90 at the distal terminal of PEG and rapidly (within a few minutes) anchored into the membranes of various cells (NIH3T3, 32D, Ba/F3, hybridoma 9E10). However, the anchored BAM90 disappeared from the cell membranes within 4-5 h in serum-free culture media, and moreover, the retention time of anchoring was shortened (1-2 h) in culture medium containing 10% FBS. We further prepared a dioleylphosphatidylethanolamine (DOPE)-PEG derivative (DOPE-BAM80, the EO unit is 80) as a double oleyl chain derivative for comparison with the single oleyl chain derivative, BAM90. The retention time of anchored DOPE-BAM80 was longer than that of BAM90 and more than 8 h in culture media with and without 10% serum. Furthermore, the treatment time of DOPE-BAM80 for anchoring was nearly as short (within a few minutes) as that of BAM90. In addition, both types of BAMs, BAM90 and DOPE-BAM80, showed no cytotoxicity. Therefore, DOPE-BAM80 is useful for protein anchoring to cells. Although the utilization of BAM90 is considered to be limited, it is expected to useful in restricted environments, for example, in tissues such as the cornea, peritoneum, bladder, and various mucosae, which are less exposed to serum. Thus, we suggest the possibility that both types of BAM can be applied to cell surface engineering.

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Section: Discussionmentioning

confidence: 99%

Rapid Protein Anchoring into the Membranes of Mammalian Cells Using Oleyl Chain and Poly(ethylene glycol) Derivatives

et al. 2004

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“…BAM contains a membrane anchoring oleyl group, a hydrophilic poly ethylene glycol (PEG) domain, and an NHS-reactive ester group (Kato et al, 2003). The BAM-coated dishes were prepared as described previously (Shimizu et al, 2012).…”

Section: Preparation Of Bam-coated Dishesmentioning

confidence: 99%

Cortical rigidity of round cells in mitotic phase and suspended state

Shimizu

Haghparast

Kihara

et al. 2012

Micron

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“…To fix non-adherent cells, we examined three types of methods which made use of a gold surface: (1) the biocompatible anchor for cell membranes (BAM: SUNBRIGHT OE-040CS) which is inserted into cell membranes without causing any damage to cells (Kato et al, 2003); (2) amino-alkanethiol (cysteamine or 8-aminoalkanethiol) that have an amino group (positive charge) with electrical affinity to connect cell membranes (negative charge); (3) dithiobis[succinimydylpropionate] (DSP) that reacts with primary amines and forms covalent amide bonds (Grubor et al, 2004).…”

Section: Introductionmentioning

confidence: 99%

Living cell positioning on the surface of gold film for SPR analysis

Yanase

Suzuki

Tsutsui

et al. 2007

Biosensors and Bioelectronics

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Living cell reactions are detected as changes of the angle of resonance (AR) for surface plasmon resonance (SPR). Since SPR reflects the events in the field of evanescence, cells need to be fixed on the sensor chip. In this study, we developed methods to fix living cells on a gold surface and to recover adherent cells from the culture dish, preserving their functions to be analyzed by SPR. Human basophils and B cells were fixed to the sensor chip by a biocompatible anchor for cell membranes (alpha-succinimidyloxysuccinyl omega-oleyloxy polyoxyethylene), aminoalkanethiol (cyteamine, 8-amino octanethiol) or an amino-reactive cross-linker (dithiobis [succinimidylpropionate]). They showed an increase of AR in response to various stimuli. RBL-2H3 cells, which firmly adhered to the culture dish, were cultured/recovered with HydroCell/simple pipetting, with RepCell/pipetting at 4 degrees C, or on normal plastic culture dishes with trypsinization or by scraping at 4 degrees C, respectively. The exocytosis of RBL-2H3 cells was largely impaired by scraping, but only slightly by the treatment with pipetting on HydroCell, on RepCell, or with trypsin. The membrane ruffling of the cells prepared by the last three treatments induced by antigens appeared the same. However, the change of AR with cells prepared by trypsin and those by scraping at 4 degrees C were lower than those by HydroCell or RepCell, suggesting that trypsin may harm molecules involved in cellular reactions. Thus, the methods of cell fixation and removal with HydroCell or RepCell should enable us to analyze various reactions in either adherent or non-adherent cells by SPR.

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Immobilized culture of nonadherent cells on an oleyl poly(ethylene glycol) ether-modified surface

Cited by 77 publications

References 7 publications

Rapid Protein Anchoring into the Membranes of Mammalian Cells Using Oleyl Chain and Poly(ethylene glycol) Derivatives

Rapid Protein Anchoring into the Membranes of Mammalian Cells Using Oleyl Chain and Poly(ethylene glycol) Derivatives

Cortical rigidity of round cells in mitotic phase and suspended state

Living cell positioning on the surface of gold film for SPR analysis

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