Emanuele Ostuni

Novartis (Switzerland)

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George M. Whitesides

Harvard University

Donald E. Ingber

Boston Children's Hospital

Shuichi Takayama

The Wallace H. Coulter Department of Biomedical Engineering

Philip R. LeDuc

Carnegie Mellon University

Robert Chapman

ARC Centre of Excellence in Advanced Molecular Imaging

Xingyu Jiang

Southern University of Science and Technology

Christopher S. Chen

Harvard University

Lin Yan

Chinese PLA General Hospital

Richard G. Weiss

Georgetown University

Michael N. Liang

University of California, Berkeley

Cooperative Institutions

Harvard University

Boston Children's Hospital

Harvard University Press

Novartis (Switzerland)

Universitat Autònoma de Barcelona

Biomedical Research Networking Center in Bioengineering, Biomaterials and Nanomedicine

Massachusetts General Hospital

Brigham and Women's Hospital

University of Oxford

Institut de Ciència de Materials de Barcelona

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Using Self-Assembled Monolayers to Pattern ECM Proteins and Cells on Substrates

Humana Press eBooks (2003)

Christopher S. Chen Emanuele Ostuni George M. Whitesides Donald E. Ingber

We present a method that uses microcontact printing of alkanethiols on gold to generate patterned substrates presenting "islands" of extracellular matrix (ECM) surrounded by nonadhesive regions such that single cells attach and spread only on the adhesive regions. We have used this micropatterning technology to demonstrate that mammalian cells can be switched between growth and apoptosis programs in the presence of saturating concentrations of growth factors by either promoting or preventing cell spreading (1). From the perspective of fundamental cell biology, these results suggested that the local differentials in growth and viability that are critical for the formation of complex tissue patterns may be generated by local changes in cell-ECM interactions. In the context of cell culture technologies, such as bioreactors and cellular engineering applications, the regulation of cell function by cell shape indicates that the adhesive microenvironment around cells can be carefully optimized by patterning a substrate in addition to using soluble factors (2). Micropatterning technology will play a central role both in our understanding how ECM and cell shape regulate cell physiology and in facilitating the development of cellular biosensor and tissue engineering applications (3-5).

Micropatterning

Microcontact Printing

10.1385/1-59259-063-2:209

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Citations (16)

<title>Soft lithography and surface chemistry: enabling tools for new bioassays</title>

Proceedings of SPIE, the International Society for Optical Engineering/Proceedings of SPIE (2002)

Bernardo D. Aumond Oliver Schueller David C. Duffy Emanuele Ostuni Enoch Kim

The process of drug discovery can be accelerated by increasing the information content of bioassays and by employing assay platforms that are amenable to high throughput screening techniques. In this paper, we demonstrate how the combination of soft lithography with controlled surface chemistry achieves these goals in a wide spectrum of bioassays. A number of soft lithographic methods can be used to generate micro-structures for the purposes of increasing assay density, diversity of test conditions and improving assay detection qualities. In addition, soft lithography, combined with specific surface chemistry modification procedures and protein engineering, may be used to control the localized molecular and biological properties of substrates, thereby enabling the development of new types of bioassays. The developed methodologies are simple, easily implemented, and lend themselves well to automation. Experimental results and prototypes are presented to illustrate the capabilities of these new techniques. For example, soft lithography and surface chemistry are employed for chemically patterning substrates, stenciling biological entities onto substrates and confining solutions. As a result, information-rich, highdensity bioassays can be obtained where biological targets, surface properties and medium solutions are carefully determined and controlled.

Soft Lithography

10.1117/12.491185

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Patterning the topographical environment for mammalian cell culture using laminar flows in capillaries

1st Annual International IEEE-EMBS Special Topic Conference on Microtechnologies in Medicine and Biology. Proceedings (Cat. No.00EX451) (2002)

Shuichi Takayama Emanuele Ostuni Xiaojie Qian J. Cooper McDonald Xingyu Jiang

This paper describes the use of patterned flows of multiple laminar streams of etching solutions in capillaries to create various topographical features with sizes of 10-200 /spl mu/m in poly(dimethylsiloxane) (PDMS). A variety of topographical features were created by using channels with obstacles, by adjusting the flow rates of etchant, or by controlling the duration of etching. Bovine capillary endothelial cells aligned parallel to features when grown inside these topographically patterned capillaries with 10 /spl mu/m ridges. The capillaries with topographical features could also be further patterned with surface-attached red blood cells and surface-adsorbed proteins, using laminar flows. This two-stage patterning produces patterns of proteins and cells on the topography with alignment between the different features and patterns. The technique allows simultaneous micropatterning of multiple cell culture environments using the same capillary system.

Micropatterning

10.1109/mmb.2000.893796

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Citations (8)

Using Self-Assembled Monolayers to Pattern ECM Proteins and Cells on Substrates

Methods in molecular biology (2009)

Emanuele Ostuni George M. Whitesides Donald E. Ingber Christopher S. Chen

Micropatterning

Microcontact Printing

10.1007/978-1-59745-413-1_12

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Citations (55)

Patterning cells and their environments using multiple laminar fluid flows in capillary networks

Proceedings of the National Academy of Sciences (1999)

Shuichi Takayama J. Cooper McDonald Emanuele Ostuni Michael N. Liang Paul J. A. Kenis

This paper describes the use of laminar flow of liquids in capillary systems to pattern the cell culture substrate, to perform patterned cell deposition, and to pattern the cell culture media. We demonstrate the patterning of the cell culture substrate with different proteins, the patterning of different types of cells adjacent to each other, the patterned delivery of chemicals to adhered cells, and performing enzymatic reactions over select cells or over a portion of a cell. This method offers a way to simultaneously control the characteristics of the surface to which cells are attached, the type of cells that are in their vicinity, and the kind of media that cells or part of a cell are exposed to. The method is experimentally simple, highly adaptable, and requires no special equipment except for an elastomeric relief that can be readily prepared by rapid prototyping.

Cell type

10.1073/pnas.96.10.5545

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Citations (516)

Nano Terra: Creating Function at Surfaces Through the Application of Chemistry and Structure. A New Business Model for Platform Technology.

Emanuele Ostuni

Nano-Terra’s technology platform is centered on the precise and localized application of chemistry and structure at the nanoand micro-scales to improve or enable function at surfaces; Professor George Whitesides of Harvard developed this technology. This technology platform is unique over other competing technologies in that it has the potential to deploy over large areas, at low cost, on curved or flexible surfaces, and at high speed. In the transition from the microscale to the nanoscale, as engineering approaches reach certain limits, chemistry and its interplay with structure are critical to achieving higher function. Such a powerful portfolio cannot be commercialized through a conventional vertically integrated business model. We have developed a collaborative model that leverages our core technical strengths to develop new products and processes with partners who manufacture and market the proposed solutions. Incentives are aligned through deal structures and co-investments. We will review our business model and our core science along with examples of how we have created value for our partners.

Microscale chemistry

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Preparation of Mixed Self-Assembled Monolayers (SAMs) That Resist Adsorption of Proteins Using the Reaction of Amines with a SAM That Presents Interchain Carboxylic Anhydride Groups

Langmuir (2000)

Robert Chapman Emanuele Ostuni Lin Yan George M. Whitesides

This paper describes a procedure for preparing mixed self-assembled monolayers (mixed SAMs) on gold that resist the nonspecific adsorption of proteins from solution. This method was tested using α-amino derivatives of ω-hydroxy- and ω-methoxy-oligo(ethylene glycols): H2N(CH2CH2O)nCH3 and H2N(CH2CH2O)nH (n = 3, 6). Mixed SAMs were prepared by allowing these amines to react with a SAM presenting interchain carboxylic anhydride groups. The resistance of the resulting surfaces to adsorption of several proteinscarbonic anhydrase (EC 4.2.1.1), ribonuclease A (EC 3.1.27.5), lysozyme (EC 3.2.1.17), and fibrinogenwas examined using surface plasmon resonance (SPR) spectroscopy. These mixed SAMs resist the nonspecific adsorption of proteins approximately as effectively as single-component SAMs prepared using the conventional method involving chemisorption of oligo(ethylene glycol)-terminated alkanethiols on gold. Characterization of the mixed SAM that presents a 1:1 mixture of −OCNH(CH2CH2O)6CH3 and CO2H/CO2- groups by polarized infrared external reflectance spectroscopy indicates that the ethylene glycol units are in an amorphous conformation. A model surface for use in studies of biospecific adsorption was synthesized by reacting the anhydride groups with a mixture of H2N(CH2CH2O)6H and H2N(CH2CH2O)6CH2CONH(CH2)6NHCOC6H4SO2NH2; the resulting system was examined for its ability to bind bovine carbonic anhydrase by SPR. The values of the relevant constants were koff = 0.0054 s-1, kon = 13 000 M-1 s-1, and Kd = 0.42 μM. These values agree with values obtained by other means. The reaction of amines with SAMs that present interchain carboxylic anhydrides provides an experimentally simple route to the formation of mixed SAMs that resist the nonspecific adsorption of proteins or that adsorb a protein of interest biospecifically.

Self-assembled monolayer

10.1021/la991579l

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Citations (251)

List of Contributors

Elsevier eBooks (2020)

Julie Allickson Rafael G. Amado Usman Azam Frank Barry Irene Aldecoa Bilbao

10.1016/b978-0-12-812034-7.01002-5

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SAMS and biofunctional surfaces. The "inert surface" problem.

George M. Whitesides Xingyu Jiang Emanuele Ostuni Robert Chapman M. Grunze

Inert

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Generation of a cellular Yin-Yang pattern

Cliff Brangwynne Kevin Kit Parker Sui Huang Emanuele Ostuni George M. Whitesides

Here we report dynamic pattern generation in a microengineered model tissue system. When two or three bovine capillary endothelial cells (BCEs) were constrained to a square adhesive island (/spl les/50/spl times/50 /spl mu/m) coated with fibronectin (FN), coordinated cell migration resulted leading to stable rotation of the entire multicellular system about its geometric center. Each cell in this dynamic pattern was led by a lamellipodium that wrapped around the trailing edge of the neighboring cell, thereby creating a static form reminiscent of the Yin-Yang (YY) symbol of the Tao religion. When single cells were constrained to these islands they exhibited "edge-seeking" behavior: the leading edge of the cell preferentially migrated along the island perimeter. This behavior stopped when the cells fully spread and assumed the square shape of the island. Similar edge-driven motility was observed in creation of the multicellular YY pattern, however, this behavior persisted. Thus, the stable YY pattern emerged from collective behavior among the different interacting cellular elements. These observations suggest that a defined adhesive field may be capable of inducing complex pattern formation in tissues by spatially coupling mechanochemical processes that mediate cell migration.

Lamellipodium

Multicellular organism

Pattern Formation

Trailing edge

10.1109/iembs.2000.897971

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