Use of photolithography to encode cell adhesive domains into protein microarrays

Ji Youn Lee, Sunny S. Shah, Christopher C. Zimmer, Gang-yu Liu, Alexander Revzin

Research output: Contribution to journalArticle

44 Citations (Scopus)

Abstract

Protein microarrays are rapidly emerging as valuable tools in creating combinatorial cell culture systems where inducers of cellular differentiation can be identified in a rapid and multiplexed fashion. In the present study, protein microarraying was combined with photoresist lithography to enable printing of extracellular matrix (ECM) protein arrays while precisely controlling "on-the-spot" cell-cell interactions. In this surface engineering approach, the micropattemed photoresist layer formed on a glass substrate served as a temporary stencil during the microarray printing, defining the micrometer-scale dimensions and the geometry of the cell-adhesion domains within the printed protein spots. After removal of the photoresist, the glass substrates contained micrometer-scale cell-adhesive regions that were encoded within 300 or 500 μm diameter protein domains. Fluorescence microscopy and atomic force microscopy (AFM) were employed to characterize protein micropatterns. When incubated with micropattemed surfaces, hepatic (HepG2) cells attached on 300 or 500 μm diameter protein spots; however, the extent of cell-cell contacts within each spot varied in accordance with dimensions of the photoresist stencil, from single cells attaching on 30 μm diameter features to multicell clusters residing on 100 or 200 μm diameter regions. Importantly, the photoresist removal process was shown to have no detrimental effects on the ability of several ECM proteins (collagens I, II, and IV and laminin) to support functional hepatic cultures. The micropatterning approach described here allows for a small cell population seeded onto a single cell culture substrate to be exposed to multiple scenarios of cell-cell and cell-surface interactions in parallel. This technology will be particularly useful for high-throughput screening of biological stimuli required for tissue specification of stem cells or for maintenance of differentiated phenotype in scarce primary cells.

Original languageEnglish (US)
Pages (from-to)2232-2239
Number of pages8
JournalLangmuir
Volume24
Issue number5
DOIs
StatePublished - Mar 4 2008

Fingerprint

Photolithography
Microarrays
photolithography
adhesives
Photoresists
Adhesives
proteins
Proteins
cells
photoresists
Extracellular Matrix Proteins
Cell culture
Printing
Substrates
Glass
Fluorescence microscopy
Cell adhesion
Laminin
Stem cells
printing

ASJC Scopus subject areas

  • Physical and Theoretical Chemistry
  • Colloid and Surface Chemistry

Cite this

Use of photolithography to encode cell adhesive domains into protein microarrays. / Lee, Ji Youn; Shah, Sunny S.; Zimmer, Christopher C.; Liu, Gang-yu; Revzin, Alexander.

In: Langmuir, Vol. 24, No. 5, 04.03.2008, p. 2232-2239.

Research output: Contribution to journalArticle

Lee, Ji Youn ; Shah, Sunny S. ; Zimmer, Christopher C. ; Liu, Gang-yu ; Revzin, Alexander. / Use of photolithography to encode cell adhesive domains into protein microarrays. In: Langmuir. 2008 ; Vol. 24, No. 5. pp. 2232-2239.
@article{0a480a18cd934fa3a04d3edee3d3882c,
title = "Use of photolithography to encode cell adhesive domains into protein microarrays",
abstract = "Protein microarrays are rapidly emerging as valuable tools in creating combinatorial cell culture systems where inducers of cellular differentiation can be identified in a rapid and multiplexed fashion. In the present study, protein microarraying was combined with photoresist lithography to enable printing of extracellular matrix (ECM) protein arrays while precisely controlling {"}on-the-spot{"} cell-cell interactions. In this surface engineering approach, the micropattemed photoresist layer formed on a glass substrate served as a temporary stencil during the microarray printing, defining the micrometer-scale dimensions and the geometry of the cell-adhesion domains within the printed protein spots. After removal of the photoresist, the glass substrates contained micrometer-scale cell-adhesive regions that were encoded within 300 or 500 μm diameter protein domains. Fluorescence microscopy and atomic force microscopy (AFM) were employed to characterize protein micropatterns. When incubated with micropattemed surfaces, hepatic (HepG2) cells attached on 300 or 500 μm diameter protein spots; however, the extent of cell-cell contacts within each spot varied in accordance with dimensions of the photoresist stencil, from single cells attaching on 30 μm diameter features to multicell clusters residing on 100 or 200 μm diameter regions. Importantly, the photoresist removal process was shown to have no detrimental effects on the ability of several ECM proteins (collagens I, II, and IV and laminin) to support functional hepatic cultures. The micropatterning approach described here allows for a small cell population seeded onto a single cell culture substrate to be exposed to multiple scenarios of cell-cell and cell-surface interactions in parallel. This technology will be particularly useful for high-throughput screening of biological stimuli required for tissue specification of stem cells or for maintenance of differentiated phenotype in scarce primary cells.",
author = "Lee, {Ji Youn} and Shah, {Sunny S.} and Zimmer, {Christopher C.} and Gang-yu Liu and Alexander Revzin",
year = "2008",
month = "3",
day = "4",
doi = "10.1021/la702883d",
language = "English (US)",
volume = "24",
pages = "2232--2239",
journal = "Langmuir",
issn = "0743-7463",
publisher = "American Chemical Society",
number = "5",

}

TY - JOUR

T1 - Use of photolithography to encode cell adhesive domains into protein microarrays

AU - Lee, Ji Youn

AU - Shah, Sunny S.

AU - Zimmer, Christopher C.

AU - Liu, Gang-yu

AU - Revzin, Alexander

PY - 2008/3/4

Y1 - 2008/3/4

N2 - Protein microarrays are rapidly emerging as valuable tools in creating combinatorial cell culture systems where inducers of cellular differentiation can be identified in a rapid and multiplexed fashion. In the present study, protein microarraying was combined with photoresist lithography to enable printing of extracellular matrix (ECM) protein arrays while precisely controlling "on-the-spot" cell-cell interactions. In this surface engineering approach, the micropattemed photoresist layer formed on a glass substrate served as a temporary stencil during the microarray printing, defining the micrometer-scale dimensions and the geometry of the cell-adhesion domains within the printed protein spots. After removal of the photoresist, the glass substrates contained micrometer-scale cell-adhesive regions that were encoded within 300 or 500 μm diameter protein domains. Fluorescence microscopy and atomic force microscopy (AFM) were employed to characterize protein micropatterns. When incubated with micropattemed surfaces, hepatic (HepG2) cells attached on 300 or 500 μm diameter protein spots; however, the extent of cell-cell contacts within each spot varied in accordance with dimensions of the photoresist stencil, from single cells attaching on 30 μm diameter features to multicell clusters residing on 100 or 200 μm diameter regions. Importantly, the photoresist removal process was shown to have no detrimental effects on the ability of several ECM proteins (collagens I, II, and IV and laminin) to support functional hepatic cultures. The micropatterning approach described here allows for a small cell population seeded onto a single cell culture substrate to be exposed to multiple scenarios of cell-cell and cell-surface interactions in parallel. This technology will be particularly useful for high-throughput screening of biological stimuli required for tissue specification of stem cells or for maintenance of differentiated phenotype in scarce primary cells.

AB - Protein microarrays are rapidly emerging as valuable tools in creating combinatorial cell culture systems where inducers of cellular differentiation can be identified in a rapid and multiplexed fashion. In the present study, protein microarraying was combined with photoresist lithography to enable printing of extracellular matrix (ECM) protein arrays while precisely controlling "on-the-spot" cell-cell interactions. In this surface engineering approach, the micropattemed photoresist layer formed on a glass substrate served as a temporary stencil during the microarray printing, defining the micrometer-scale dimensions and the geometry of the cell-adhesion domains within the printed protein spots. After removal of the photoresist, the glass substrates contained micrometer-scale cell-adhesive regions that were encoded within 300 or 500 μm diameter protein domains. Fluorescence microscopy and atomic force microscopy (AFM) were employed to characterize protein micropatterns. When incubated with micropattemed surfaces, hepatic (HepG2) cells attached on 300 or 500 μm diameter protein spots; however, the extent of cell-cell contacts within each spot varied in accordance with dimensions of the photoresist stencil, from single cells attaching on 30 μm diameter features to multicell clusters residing on 100 or 200 μm diameter regions. Importantly, the photoresist removal process was shown to have no detrimental effects on the ability of several ECM proteins (collagens I, II, and IV and laminin) to support functional hepatic cultures. The micropatterning approach described here allows for a small cell population seeded onto a single cell culture substrate to be exposed to multiple scenarios of cell-cell and cell-surface interactions in parallel. This technology will be particularly useful for high-throughput screening of biological stimuli required for tissue specification of stem cells or for maintenance of differentiated phenotype in scarce primary cells.

UR - http://www.scopus.com/inward/record.url?scp=41849092997&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=41849092997&partnerID=8YFLogxK

U2 - 10.1021/la702883d

DO - 10.1021/la702883d

M3 - Article

C2 - 18198912

AN - SCOPUS:41849092997

VL - 24

SP - 2232

EP - 2239

JO - Langmuir

JF - Langmuir

SN - 0743-7463

IS - 5

ER -