Arraying heterotypic single cells on photoactivatable cell-culturing substrates

Yukiko Kikuchi, Jun Nakanishi, Takahiro Shimizu, Hidekazu Nakayama, Satoshi Inoue, Kazuo Yamaguchi, Hideo Iwai, Yasuhiko Yoshida, Yasuhiro Horiike, Tohru Takarada, Mizuo Maeda

研究成果: Article

46 引用 (Scopus)

抄録

This article describes a photochemical method for the site-selective assembly of heterotypic cells on a glass substrate modified with a silane coupling agent having a caged functional group. Silane coupling agents having a carboxyl (COOH), amino (NH2), hydroxyl (OH), or thiol (SH) group protected by a photocleavable 2-nitrobenzyl group were synthesized to modify the surfaces of glass coverslips. The caged substrates were first coated by the adsorption of a blocking agent, bovine serum albumin (BSA), to make the entire surface non-cell-adhesive and then irradiated at 365 nm under a standard fluorescence microscope. The photocleavage reaction on the surface was followed by contact angle measurements and X-ray photoelectron spectroscopy. When COS7, NIH3T3, and HEK293 cells were seeded onto these substrates in a serum-free medium, the cells adhered selectively and efficiently to the irradiated regions on the caged NH2 substrate, whereas the other caged COOH, SH, and OH substrates were nonphotoactivatable for cell adhesion. Qualitative and quantitative analysis of BSA adsorbed to the uncaged substrates revealed that this highly efficient photoactivation on the caged NH2 substrate arose because of the following reasons: (i) upon photoactivation, BSA adsorbed in advance on the 2-nitrobenzyl groups was readsorbed onto the uncaged functional groups and (ii) BSA readsorbed onto the NH2 groups became unable to passivate the surface against cell adhesion whereas BSA on the other groups still had normal passivating activity. It was also demonstrated that heterotypic single COS7, NIH3T3, and HEK293 cells were positioned at any desired arrangement on the caged NH2 substrate by repeating the UV irradiation at optimized array spot sizes and cell seeding in optimized cell concentrations. The present method will be particularly useful in studying the dynamic processes of cell-cell interactions at a single-cell level.

元の言語English
ページ(範囲)13084-13095
ページ数12
ジャーナルLangmuir
24
発行部数22
DOI
出版物ステータスPublished - 2008 11 18
外部発表Yes

Fingerprint

cell culturing
Bovine Serum Albumin
Substrates
serums
cells
albumins
Silanes
Coupling agents
Cell adhesion
Functional groups
silanes
Glass
adhesion
Serum-Free Culture Media
Angle measurement
Sulfhydryl Compounds
Hydroxyl Radical
Contact angle
glass
Adhesives

ASJC Scopus subject areas

  • Materials Science(all)
  • Condensed Matter Physics
  • Surfaces and Interfaces
  • Spectroscopy
  • Electrochemistry

これを引用

Kikuchi, Y., Nakanishi, J., Shimizu, T., Nakayama, H., Inoue, S., Yamaguchi, K., ... Maeda, M. (2008). Arraying heterotypic single cells on photoactivatable cell-culturing substrates. Langmuir, 24(22), 13084-13095. https://doi.org/10.1021/la8024414

Arraying heterotypic single cells on photoactivatable cell-culturing substrates. / Kikuchi, Yukiko; Nakanishi, Jun; Shimizu, Takahiro; Nakayama, Hidekazu; Inoue, Satoshi; Yamaguchi, Kazuo; Iwai, Hideo; Yoshida, Yasuhiko; Horiike, Yasuhiro; Takarada, Tohru; Maeda, Mizuo.

:: Langmuir, 巻 24, 番号 22, 18.11.2008, p. 13084-13095.

研究成果: Article

Kikuchi, Y, Nakanishi, J, Shimizu, T, Nakayama, H, Inoue, S, Yamaguchi, K, Iwai, H, Yoshida, Y, Horiike, Y, Takarada, T & Maeda, M 2008, 'Arraying heterotypic single cells on photoactivatable cell-culturing substrates', Langmuir, 巻. 24, 番号 22, pp. 13084-13095. https://doi.org/10.1021/la8024414
Kikuchi Y, Nakanishi J, Shimizu T, Nakayama H, Inoue S, Yamaguchi K その他. Arraying heterotypic single cells on photoactivatable cell-culturing substrates. Langmuir. 2008 11 18;24(22):13084-13095. https://doi.org/10.1021/la8024414
Kikuchi, Yukiko ; Nakanishi, Jun ; Shimizu, Takahiro ; Nakayama, Hidekazu ; Inoue, Satoshi ; Yamaguchi, Kazuo ; Iwai, Hideo ; Yoshida, Yasuhiko ; Horiike, Yasuhiro ; Takarada, Tohru ; Maeda, Mizuo. / Arraying heterotypic single cells on photoactivatable cell-culturing substrates. :: Langmuir. 2008 ; 巻 24, 番号 22. pp. 13084-13095.
@article{7db38a8509f1468297dbd9bfea4337fb,
title = "Arraying heterotypic single cells on photoactivatable cell-culturing substrates",
abstract = "This article describes a photochemical method for the site-selective assembly of heterotypic cells on a glass substrate modified with a silane coupling agent having a caged functional group. Silane coupling agents having a carboxyl (COOH), amino (NH2), hydroxyl (OH), or thiol (SH) group protected by a photocleavable 2-nitrobenzyl group were synthesized to modify the surfaces of glass coverslips. The caged substrates were first coated by the adsorption of a blocking agent, bovine serum albumin (BSA), to make the entire surface non-cell-adhesive and then irradiated at 365 nm under a standard fluorescence microscope. The photocleavage reaction on the surface was followed by contact angle measurements and X-ray photoelectron spectroscopy. When COS7, NIH3T3, and HEK293 cells were seeded onto these substrates in a serum-free medium, the cells adhered selectively and efficiently to the irradiated regions on the caged NH2 substrate, whereas the other caged COOH, SH, and OH substrates were nonphotoactivatable for cell adhesion. Qualitative and quantitative analysis of BSA adsorbed to the uncaged substrates revealed that this highly efficient photoactivation on the caged NH2 substrate arose because of the following reasons: (i) upon photoactivation, BSA adsorbed in advance on the 2-nitrobenzyl groups was readsorbed onto the uncaged functional groups and (ii) BSA readsorbed onto the NH2 groups became unable to passivate the surface against cell adhesion whereas BSA on the other groups still had normal passivating activity. It was also demonstrated that heterotypic single COS7, NIH3T3, and HEK293 cells were positioned at any desired arrangement on the caged NH2 substrate by repeating the UV irradiation at optimized array spot sizes and cell seeding in optimized cell concentrations. The present method will be particularly useful in studying the dynamic processes of cell-cell interactions at a single-cell level.",
author = "Yukiko Kikuchi and Jun Nakanishi and Takahiro Shimizu and Hidekazu Nakayama and Satoshi Inoue and Kazuo Yamaguchi and Hideo Iwai and Yasuhiko Yoshida and Yasuhiro Horiike and Tohru Takarada and Mizuo Maeda",
year = "2008",
month = "11",
day = "18",
doi = "10.1021/la8024414",
language = "English",
volume = "24",
pages = "13084--13095",
journal = "Langmuir",
issn = "0743-7463",
publisher = "American Chemical Society",
number = "22",

}

TY - JOUR

T1 - Arraying heterotypic single cells on photoactivatable cell-culturing substrates

AU - Kikuchi, Yukiko

AU - Nakanishi, Jun

AU - Shimizu, Takahiro

AU - Nakayama, Hidekazu

AU - Inoue, Satoshi

AU - Yamaguchi, Kazuo

AU - Iwai, Hideo

AU - Yoshida, Yasuhiko

AU - Horiike, Yasuhiro

AU - Takarada, Tohru

AU - Maeda, Mizuo

PY - 2008/11/18

Y1 - 2008/11/18

N2 - This article describes a photochemical method for the site-selective assembly of heterotypic cells on a glass substrate modified with a silane coupling agent having a caged functional group. Silane coupling agents having a carboxyl (COOH), amino (NH2), hydroxyl (OH), or thiol (SH) group protected by a photocleavable 2-nitrobenzyl group were synthesized to modify the surfaces of glass coverslips. The caged substrates were first coated by the adsorption of a blocking agent, bovine serum albumin (BSA), to make the entire surface non-cell-adhesive and then irradiated at 365 nm under a standard fluorescence microscope. The photocleavage reaction on the surface was followed by contact angle measurements and X-ray photoelectron spectroscopy. When COS7, NIH3T3, and HEK293 cells were seeded onto these substrates in a serum-free medium, the cells adhered selectively and efficiently to the irradiated regions on the caged NH2 substrate, whereas the other caged COOH, SH, and OH substrates were nonphotoactivatable for cell adhesion. Qualitative and quantitative analysis of BSA adsorbed to the uncaged substrates revealed that this highly efficient photoactivation on the caged NH2 substrate arose because of the following reasons: (i) upon photoactivation, BSA adsorbed in advance on the 2-nitrobenzyl groups was readsorbed onto the uncaged functional groups and (ii) BSA readsorbed onto the NH2 groups became unable to passivate the surface against cell adhesion whereas BSA on the other groups still had normal passivating activity. It was also demonstrated that heterotypic single COS7, NIH3T3, and HEK293 cells were positioned at any desired arrangement on the caged NH2 substrate by repeating the UV irradiation at optimized array spot sizes and cell seeding in optimized cell concentrations. The present method will be particularly useful in studying the dynamic processes of cell-cell interactions at a single-cell level.

AB - This article describes a photochemical method for the site-selective assembly of heterotypic cells on a glass substrate modified with a silane coupling agent having a caged functional group. Silane coupling agents having a carboxyl (COOH), amino (NH2), hydroxyl (OH), or thiol (SH) group protected by a photocleavable 2-nitrobenzyl group were synthesized to modify the surfaces of glass coverslips. The caged substrates were first coated by the adsorption of a blocking agent, bovine serum albumin (BSA), to make the entire surface non-cell-adhesive and then irradiated at 365 nm under a standard fluorescence microscope. The photocleavage reaction on the surface was followed by contact angle measurements and X-ray photoelectron spectroscopy. When COS7, NIH3T3, and HEK293 cells were seeded onto these substrates in a serum-free medium, the cells adhered selectively and efficiently to the irradiated regions on the caged NH2 substrate, whereas the other caged COOH, SH, and OH substrates were nonphotoactivatable for cell adhesion. Qualitative and quantitative analysis of BSA adsorbed to the uncaged substrates revealed that this highly efficient photoactivation on the caged NH2 substrate arose because of the following reasons: (i) upon photoactivation, BSA adsorbed in advance on the 2-nitrobenzyl groups was readsorbed onto the uncaged functional groups and (ii) BSA readsorbed onto the NH2 groups became unable to passivate the surface against cell adhesion whereas BSA on the other groups still had normal passivating activity. It was also demonstrated that heterotypic single COS7, NIH3T3, and HEK293 cells were positioned at any desired arrangement on the caged NH2 substrate by repeating the UV irradiation at optimized array spot sizes and cell seeding in optimized cell concentrations. The present method will be particularly useful in studying the dynamic processes of cell-cell interactions at a single-cell level.

UR - http://www.scopus.com/inward/record.url?scp=57349098402&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=57349098402&partnerID=8YFLogxK

U2 - 10.1021/la8024414

DO - 10.1021/la8024414

M3 - Article

C2 - 18925763

AN - SCOPUS:57349098402

VL - 24

SP - 13084

EP - 13095

JO - Langmuir

JF - Langmuir

SN - 0743-7463

IS - 22

ER -