WO2013156941A1 - Dispositif permettant d'obtenir des cultures cellulaires tridimensionnelles, procédé pour sa mise en œuvre, et utilisation d'un tel dispositif - Google Patents

Dispositif permettant d'obtenir des cultures cellulaires tridimensionnelles, procédé pour sa mise en œuvre, et utilisation d'un tel dispositif Download PDF

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Publication number
WO2013156941A1
WO2013156941A1 PCT/IB2013/053041 IB2013053041W WO2013156941A1 WO 2013156941 A1 WO2013156941 A1 WO 2013156941A1 IB 2013053041 W IB2013053041 W IB 2013053041W WO 2013156941 A1 WO2013156941 A1 WO 2013156941A1
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WIPO (PCT)
Prior art keywords
micro
substrate
structures
cells
biocompatible
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PCT/IB2013/053041
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English (en)
Inventor
Enzo Di Fabrizio
Tania LIMONGI
Francesco Gentile
Roberto MAROTTA
Fabio Benfenati
Fabrizia CESCA
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Fondazione Istituto Italiano Di Tecnologia
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    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
    • C12M21/00—Bioreactors or fermenters specially adapted for specific uses
    • C12M21/18—Apparatus specially designed for the use of free, immobilized or carrier-bound enzymes
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
    • C12M23/00—Constructional details, e.g. recesses, hinges
    • C12M23/20—Material Coatings
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
    • C12M23/00—Constructional details, e.g. recesses, hinges
    • C12M23/34—Internal compartments or partitions

Definitions

  • the present invention relates to a device for obtaining three-dimensional cell cultures, to a method for the implementation thereof, and to a use of such device for obtaining suspended cells not adherent to the substrate.
  • Propensity of a cell to adhere, grow, and differentiate on a compact substrate (or support) or on a scaffold having a supporting function is one of the most important aspect of tissue engineering and biomaterials characterization.
  • the ratio between cells and substrate is controlled by the geometrical characteristics and the chemical and physical properties of the support, with the aim of producing cell cultures in vitro with an organization and functionality as near to those of native tissues as possible.
  • the ability to grow by responding to the growth support topographic characteristics is well known for different cell types; they show interesting growth responses when plated on substrates characterized in surface by different geometries and different degrees of superhydrofobicity.
  • Document WO 2010/023635 discloses a device in which, on a substrate, a plurality of micro-structures mutually spaced apart in a periodical manner are obtained, so as to make the substrate super hydrophobic, intended to the use for the concentration and localization of a solute in a reduced portion of space.
  • matrices of synthetic or natural biodegradable polymers there are matrices of synthetic or natural biodegradable polymers.
  • polymers with different porosities can be obtained in devices such as membranes, films, and nano-tubes, as described in Langer, R. & Tirrell, D. A. Designing materials for biology and medicine. Nature 428, 487-492, doi: 10.1038/nature02388 (2004).
  • tubes made of a collagen matrix are used to promote nervous tissue regeneration, and materials in gel form are used mainly for isolation or as substrates for the connective tissue regeneration.
  • these devices are composed of a porous matrix on which the cells attach. Within the pores, in addition to the cells, the fluid acting as a nutrient also must fit in.
  • Cell growth is strongly related to the amount of nutrients that are present in the growth medium and the motion type of the fluid. When a uniform culture inside the whole cellular construct is required, it will be necessary that the nutrient fluid succeeds in invading each pore of the matrix. On the other hand, the cell growth, in time, fills the voids that are present in the matrix, thus causing the nutrient flow to decrease.
  • the known supporting devices for the three-dimensional growth of suspended cells are not perfectly able to provide a sufficient amount of nutrients, while suitably removing waste products.
  • object of the present invention is to propose a device for obtaining three- dimensional cell cultures, which is capable of overcoming the above-mentioned problems, and allowing the growth of suspended cells also for cell types that usually grow while being adherent to the substrate, particularly for nervous tissue regeneration applications.
  • a further object of the present invention is to propose a method for manufacturing the above-mentioned device and a use of such device for obtaining suspended cells not adherent to the substrate.
  • FIG. 1 is a top view of a device according to the invention
  • Fig. 2 is a perspective enlargement of one of the nano-structures of the device of Fig. 1, and
  • Fig. 3 is a side view of the nano-structure of Fig. 2.
  • the invention uses techniques of micro-fabrication for manufacturing a super hydrophobic device having structures with side walls at least partially nano-patterned, used in cell culture methods for obtaining cells that are suspended, or not completely adherent to the substrate, in particular for the three-dimensional growth of hippocampal neuronal cells and for the growth of hippocampal neurons and glial cells co-culture.
  • Such device combines two types of topographic details, having dimensions on a different scale, respectively, in particular micrometric structures conferring superhydrofobicity to the substrate surface, while nano-metric features of such micrometric structures make the stratification in three dimensions of the cell cultures possible.
  • the surface geometry of the supports is characterized by large free spaces between a micro-structure and the adjacent one, and this allows a better recirculation of the nutrient substances and easier gaseous exchanges between the cells and the growth medium.
  • the device of the present invention allows achieving results that are qualitatively higher by providing to the cells the correct amount of nutrients and mechanical stimuli for the development of co-cultures and tissues for biomedical applications that are more reproducible and cost effective.
  • the invention aims to manufacturing nano-patterned superhydrofobic devices for the three-dimensional growth of normal cells (also in co-culture), for the study of the spatial arrangement of tumoral cell cultures; for pharmacological and toxicological screening tests in vitro, aimed to cell therapy.
  • the invention allows performing tissue engineering applications, such as nervous regeneration.
  • the device according to the invention comprising a substrate 1 for example made of silicon or a biocompatible or bioerodible polymeric material, and a plurality of micro-structures 2, in particular micro-cylinders, extending from the substrate.
  • a substrate 1 for example made of silicon or a biocompatible or bioerodible polymeric material
  • micro-structures 2 can advantageously have a section in a circular, squared, rectangular, hexagonal shape, or any other shape.
  • Such micro-structures 2 are preferably arranged orthogonally to the surface of the substrate 1 and are located on the substrate 1 according to a periodical grid so as to make the substrate 1 super hydrophobic.
  • the term super hydrophobic in the present description is meant a surface on which the contact angle that a drop of (distilled) water forms with the surface itself is above 150°.
  • the contact angle is the angle formed by the tangent at the liquid-vapor interface with the tangent at the liquid-solid interface.
  • Obtaining micro-patterned super hydrophobic surfaces is a function of the surface density and the transversal dimension of the micro-structures.
  • the design principles for manufacturing the above-mentioned micro-patterned super hydrophobic surfaces are per se known and are described, for example, in de Angelis, F. et al. "Breaking the diffusion limit with super-hydrophobic delivery of molecules to plasmonic nanofocusing SERS structures", Nature Photonics, Volume 5, Issue 1 1, pp. 682-687 (201 1), incorporated herein for effect of such quote.
  • the substrate 1 has a resistivity of about 5-10 ⁇ / ⁇ .
  • micro-structures 2 are obtained by lithography and etching processes, or micromoulding processes, per se known, as described herein below.
  • the micro-structures 2 are spaced apart on the substrate 1 by a predetermined distance 4 preferably ranging within 15-30 ⁇ .
  • the micro-structures 2 have preferably a base width ranging within 5-20 ⁇ , and a height ranging within 5-30 ⁇ .
  • base width is meant in the present description the minimum dimension among those defining the section shape.
  • the base width and the distance 4 between the micro-structures 2 are selected according to a per se known criterion from the above-mentioned article.
  • the device In order to use the device for the objects that are the subject-matter of the invention, it is preferred to apply on the device surface coatings suitable to promote the growth of cell cultures while maintaining or imposing the superhydrofobicity properties.
  • a biocompatible coating (where the substrate is not per se biocompatible) can be contemplated, for example, a metallic, glass, or glass-ceramic coating, and an optional adhesion layer for said biocompatible coating, applied on the substrate surface.
  • a thin hydrophobic polymeric coating for example, a fluorinated polymer, as obtained by the polymerisation of octafluorocyclobutane, can finally be applied to the biocompatible coating, having thicknesses such as not to change the device superficial biocompatibility.
  • FIG. 2 a perspective enlargement is illustrated of one of the micro-structures of the device of Fig. 1.
  • the side walls of the micro-structure 2 are not smooth, but they are patterned, i.e., they have alternated projections 14 and recesses 16, with a predetermined pitch (or spatial distance) depending on the duration of the plasma isotropic dry etching step and the deposition step of a passivation layer, described herein below.
  • the height of the projections 14 or recesses 16 can be constant along the whole wall of a micro-structure, or it can vary, as well as the height between adjacent projections 14 and recesses 16 can be the same or different.
  • smooth is meant that the side wall of a micro- structure, as seen in section of a plane parallel to the main extension axis of the micro- structure, has a rectilinear profile r and it does not have projections and recesses.
  • FIG. 3 a side view of the micro-structure 2 of Fig. 2 is shown, in which the height h of the micro-structure 2 and the height i separating two adjacent projections 14 are highlighted, respectively.
  • the micro-structures 2 do not have completely patterned side walls, but they have an alternation of patterned zones (bands) and smooth zones (bands) without projections 14 and recesses 16.
  • the device of the present invention also comprises a plurality of electrodes, each associated to a micro-structure 2 (whether it is of the type with completely patterned side walls, or of the type with side walls with alternation of smooth zones and patterned zones), connected to conductive tracks manufactured in a per se known manner in the substrate 1. In such a manner, it is possible to apply voltages to such electrodes, therefore to such micro-structures 2, thus obtaining an electric device.
  • the device of the present invention also comprises a plurality of micro- channels, which can be manufactured in a per se known manner, allowing controlled changes of medium or cytotoxicity tests.
  • the device of the present invention comprises an alphanumerical grid, located in the peripheral zone of the substrate 1 , so as to easily allow counting and localizing the cells, to allow an analysis by different techniques, such as confocal optical microscope, or atomic force microscope, Raman spectroscopy, or also SEM.
  • the substrate is made of silicon or a bioerodible and biocompatible polymeric material.
  • the substrate 1 is made of silicon, it is first cleaned in a per se known manner.
  • a per se known optic lithography step is performed, by using an optical mask obtained by a laser writing process.
  • a regular pattern of disks is obtained within a covering layer that had previously deposited by spin coating on the cleaned substrate 1.
  • a covering mask comprising a plurality of disks or polygons by carrying out an optic lithography on a uniform covering layer deposited on the substrate 1, for example, a resist, using in a known manner a light beam having a predetermined wavelength ⁇ ranging within between 350 and 450nm.
  • the covering layer externally to the disks (or polygons) is removed, and in the following step a dry etching step of the DRIE type is carried out, of the substrate 1 , by using the pre-prepared covering mask.
  • the above-described micro-structures 2 are obtained (thus having all the spatial disposition characteristics illustrated above) and having an aspect ratio, i.e., the ratio between the height of the micro-structure 2 and the area of the base thereof, higher than 2.
  • micro-structures 2 are obtained, having nano- patterned side walls as shown in Fig. 2.
  • any deviation from r would unbalance the process: if tj/t 2 is higher than r, passivation step would dominate over dry etching, and therefore the micro-structures 2 would be frustoconical shaped; if t t /t 2 is lesser than r, the dry etching would dominate over the passivation step, and the micro-structures 2 would be shaped into an upside-down frustum.
  • the covering layer is removed in a per se known manner with a removal solution. Finally, the above-described coatings are deposited on the substrate.
  • the substrate 1 of the device of the invention is made of a bioerodible and biocompatible polymeric material
  • a mould (master) of a rigid and undeformable material is made, such as, for example, silicon.
  • the mould is manufactured starting from a starting substrate, in a similar manner to what has been described above with reference to the implementations steps of the micro-structures 2.
  • the parameters d, I, and / remain the same and are inspired to the same designing criteria.
  • the mould manufacturing process provides for the use of the same micro-optic lithography techniques as described above. Particularly, the same covering layer is used to obtain a covering mask complementary to that described above.
  • a uniform covering layer is deposited, by spin coating, on which, in a per se known manner by a lithographic process, a regular pattern of disks (or other polygons) is obtained.
  • a covering mask is thus obtained.
  • the covering layer within the disks (or other polygons) is removed in a known manner, and, in the following step, a dry etching step of the DRIE type is carried out, as described above, by virtue of which the starting substrate is eroded at the disks, thus obtaining a mould comprising a plurality of holes having the same characteristics of the micro-structures 2 described above.
  • such holes by virtue of the alternation of steps of the DRIE process, have nano-patterned side walls.
  • the covering mask is removed in a known manner and the mould is then further cleaned.
  • the so-obtained mould is silanized in a vacuum reaction bell, and it is subsequently "baked” at temperatures ranging between 100° C and 120° C.
  • bioerodible and biocompatible polymeric material is brought to melt in a known manner on a support.
  • the mould is placed in contact with said molten bioerodible and biocompatible polymeric material, avoiding the formation of air bubbles at the interface between the mould and the biocompatible material, so as to carry out a micromoulding process as described herein.
  • a forming or moulding step of the bioerodible and biocompatible polymeric material layer is carried out in a known manner.
  • the forming or moulding process comprises two steps:
  • bioerodible and biocompatible polymeric material is released from the mould by first removing the support, then separating the mould and the biocompatible material substrate with a slight pulling action.
  • a first implementation example of the present invention comprises the use of a P doped silicon substrate 1 and the micro-structures 2 are arranged on the substrate 1 according to a periodical hexagonal grid and are spaced apart by a distance 4 of 20 ⁇ .
  • the micro- structures 2 have a base width of ⁇ and a height of ⁇ .
  • the micro-structure 2, having a P doped silicon core 6, are coated with a first titanium layer 8, completely covering the core 6, a second gold layer 10, covering the first layer 8, and a third layer 12, covering the second layer 10, of a hydrophobic polymer obtained by performing a polymerisation of octafluorocyclobutane (C 4 Fg).
  • the silicon substrate is first cleaned with acetone and isopropanol, and, after immersion in a 4% hydrofluoric acid solution, it is cleaned with deionized water and dried with nitrogen.
  • a Mask Aligner of the type MA6/BA6, SUSS MICROTEC is used, obtaining a regular pattern of disks (or other polygon) within a covering layer of AZ5214 negative resist.
  • the Mask Aligner produce a resist mask by carrying out an optic lithography on a uniform resist layer deposited on the substrate 1 , using in a known manner a light beam having a wavelength ⁇ ranging between 350 and 450nm.
  • the dry etching step is carried out with a ICP-RIE, SI 500, Sentech Instrument GmbH instrument, and the used DRIE process repeatedly alternates three steps:
  • a first titanium layer having a thickness of 5nm and a second gold layer having a thickness of lOOnm are deposited on the substrate 1 by sputtering.
  • the titanium layer improves adhesion of the gold layer.
  • the second layer is coated with a 5nm third layer of a hydrophobic polymer obtained by polymerisation of octafluorocyclobutane (C 4 F 8 ).
  • a second implementation example of the present invention comprises the use of a substrate 1 of polycaprolactone (PLC), and the micro-structures 2 are arranged on the substrate 1 according to a periodical hexagonal grid and are spaced apart by a distance 4 of 20 ⁇ .
  • the micro-structures 2 have a base width of ⁇ and a height of ⁇ .
  • a silicon master is obtained by using the same micro-optic lithography techniques described above. Particularly, negative resist layer of the AZ5214 type is used to obtain a covering mask complementary to that described above.
  • the covering mask is removed by sonication in acetone at 55° C, and the mould is cleaned by a piranha solution.
  • the so-obtained mould is silanized in a vacuum reaction bell, and it is then "baked" at 112° C.
  • the forming or moulding process comprises the steps of:
  • a drop of growth medium of the type described herein below, is deposited on the surface of the device of the present invention, it keeps an almost spherical shape with a contact angle at the air- water interface that can be theoretically predicted in a per se known manner, by using the Cassie and Baxter model. According to such model, the wettability behaviour of the device surface is described by the sole parameter f. When f tends to zero, at the interface with the device substrate 1, the liquid mostly "senses" air, and the drop would resemble a perfect sphere.
  • the device can be suitably used to manipulate different samples of biomedical interest.
  • Ii is generally known that the hydro-repellent properties of the superhydrofobic materials reduce the water contact area on their surface, thereby minimizing the absortion of particles or molecules.
  • the nano-patterned superhydrofobic surface of the device of the invention is used as a substrate for cell cultures.
  • the nano-patterned super hydrophobic device (DSN) of the present invention such device is coated with a material promoting the adhesion for the cells to be grown, such as, for example poly-D-lysine.
  • a material promoting the adhesion for the cells to be grown such as, for example poly-D-lysine.
  • cells to be grown are deposited on the device, preferably, a predetermined number of such cells is deposited in a single drop of a predetermined growth medium (Plating Medium -PM).
  • the drop deposited on the DSN surface takes an almost spherical shape, with a contact angle above 160°, which is maintained by the relative humidity that is present in the incubator of the cell culture.
  • the high relative humidity rate prevents a rapid PM evaporation and a reduction in the diameter of the medium drop in which the cells are dispersed.
  • the cells are left in incubation in superhydrofobicity for a predetermined time interval ranging between 4 and 6 hours, and then the PM is removed and replaced with maintenance medium.
  • the cells build up in a very localized region, increasing the density thereof, strengthening cell adhesion and their ability to form extensions forming points extending from a micro-structure 2 to an adjacent one.
  • An example of a method for obtaining three-dimensional cell cultures of suspended cells is described herein below, such an example being given by way of illustrative example, non- limiting to the scope of the invention.
  • a device having an area of 1cm 2 is coated with poly-D-lysine (in a concentration of 1- 10( ⁇ g/ml). Subsequently, 50,000 hippocampal cells are deposited on the device in a single drop of ⁇ PM containing 10% horse serum and 2mM glutamine and antibiotics. The drop takes an almost spherical shape, with a contact angle above 160° kept by a relative humidity of more than 80% that is present in the incubator of the cell culture at 37° C.
  • the device of the present invention is particularly suitable for nervous regeneration applications, when manufactured with biocompatible and biodegradable materials. Particularly, it can act as an aid by combining the principles of:
  • a vertically patterned silicon device with micro-structures 2 aligned and periodically spaced apart is able to control wettability and have a distinctive role in the three-dimensional culture of cells.
  • This new type of three-dimensional growth device can be easily seeded by an automatic process, making its high-throughput use possible for any laboratory.

Abstract

L'invention concerne un dispositif permettant d'obtenir des cultures cellulaires tridimensionnelles, comprenant : un substrat (1) ; une pluralité de microstructures (2) qui dépassent de la surface du substrat (1) et sont disposées sur un tel substrat (1) selon une grille périodique de façon à rendre le substrat (1) super-hydrophobe. Le dispositif est caractérisé en ce que les microstructures (2) ont des parois latérales à motifs au moins partiellement nanométriques et ont des protubérances (14) et des cavités (16) en alternance à une distance prédéfinie.
PCT/IB2013/053041 2012-04-17 2013-04-17 Dispositif permettant d'obtenir des cultures cellulaires tridimensionnelles, procédé pour sa mise en œuvre, et utilisation d'un tel dispositif WO2013156941A1 (fr)

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ITTO2012A000331 2012-04-17
IT000331A ITTO20120331A1 (it) 2012-04-17 2012-04-17 Dispositivo per l'ottenimento di colture cellulari in tre dimensioni, procedimento per la sua realizzazione e impiego di tale dispositivo

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Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US11326138B2 (en) 2017-05-01 2022-05-10 University Of Kentucky Research Foundation Cell culture device and methods of use thereof
US11518971B2 (en) 2018-11-27 2022-12-06 Research Triangle Institute Method and apparatus for spatial control of cellular growth

Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2005005679A2 (fr) * 2003-04-28 2005-01-20 Nanosys, Inc. Surfaces superhydrophobes, methodes de leur construction et leurs utilisations
WO2010023635A1 (fr) 2008-08-29 2010-03-04 Calmed S.R.L. Dispositif de concentration et de localisation d’un soluté et procédé de concentration et de localisation d’un soluté
DE102009037011B3 (de) * 2009-08-08 2011-05-12 Helmholtz-Zentrum Berlin Für Materialien Und Energie Gmbh Molekulares Lithographieverfahren
US20110250376A1 (en) * 2010-04-07 2011-10-13 California Institute Of Technology Simple method for producing superhydrophobic carbon nanotube array

Patent Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2005005679A2 (fr) * 2003-04-28 2005-01-20 Nanosys, Inc. Surfaces superhydrophobes, methodes de leur construction et leurs utilisations
WO2010023635A1 (fr) 2008-08-29 2010-03-04 Calmed S.R.L. Dispositif de concentration et de localisation d’un soluté et procédé de concentration et de localisation d’un soluté
DE102009037011B3 (de) * 2009-08-08 2011-05-12 Helmholtz-Zentrum Berlin Für Materialien Und Energie Gmbh Molekulares Lithographieverfahren
US20110250376A1 (en) * 2010-04-07 2011-10-13 California Institute Of Technology Simple method for producing superhydrophobic carbon nanotube array

Non-Patent Citations (8)

* Cited by examiner, † Cited by third party
Title
A. RANELLA; M. BARBEROGLOU: "Tuning cell adhesion by controlling the roughness and wettability of 3D micro(nano silicon structures", ACTA BIOMATERIALIA, vol. 6, 18 January 2010 (2010-01-18), pages 2711 - 2720, XP002691330 *
DE ANGELIS. F. ET AL.: "Breaking the diffusion limit with super-hydrophobic delivery of molecules to plasmonic nanofocusing SERS structures", NATURE PHOTONICS, vol. 5, no. 11, 2011, pages 682 - 687
LANGER, R.; TIRRELL, D. A: "Designing materials for biology and medicine", NATURE, vol. 428, 2004, pages 487 - 492
MANHUI SUN ET AL: "Artificial Lotus Leaf by Nanocasting", vol. 21, no. 19, 1 January 2005 (2005-01-01) - 1 January 2005 (2005-01-01), pages 8978 - 8981, XP002667652, ISSN: 0743-7463, Retrieved from the Internet <URL:http://pubs.acs.org/doi/abs/10.1021/la050316q> [retrieved on 20050816], DOI: 10.1021/LA050316Q *
ROACH P ET AL: "Progress in superhydrophobic surface development", 1 January 2008, SOFT MATTER, R S C PUBLICATIONS, GB, PAGE(S) 224 - 240, ISSN: 1744-683X, XP002558612 *
ROSA DI MUNDO; ROBERTO GRISTINA: "Mcro-/Nanoscale Structuring of cell culture substrates with fluorocarbon plasmas", PLASMA, PROCESSES AND POLYMERS, vol. 7, 27 September 2009 (2009-09-27), pages 212 - 223, XP002691331 *
TAKAHIRO ISHIZAKI; NAGAHIRO SAITO: "Correlation of cell adhesive behaviour on superhydrophobic, superhydrophilic, and micropatternd superhydrophobic/superhydrophilic surfaces in their surface chemistry", LANGMUIR ARTICLE, vol. 26, no. 11, 2 April 2010 (2010-04-02), pages 8147 - 8154, XP002691332 *
TAN, J. L. ET AL.: "Cells lying on a bed of microneedles: An approach to isolate mechanical force", PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, vol. 100, 2003, pages 1484 - 1489

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US11326138B2 (en) 2017-05-01 2022-05-10 University Of Kentucky Research Foundation Cell culture device and methods of use thereof
US11518971B2 (en) 2018-11-27 2022-12-06 Research Triangle Institute Method and apparatus for spatial control of cellular growth

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