Cell Co-culture Patterning Using Aqueous Two-phase Systems

Frampton, John P.; White, Joshua B.; Abraham, Abin T.; Takayama, Shuichi

doi:10.3791/50304

Cited by 28 publications

(43 citation statements)

References 29 publications

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“…Although not tested here, based upon the results of this and previous studies [11–14,21–24], it is reasonable to expect that this method can be extended for use with a variety of eukaryotic cell types and bacteria, both pathogenic and commensal ones.…”

Section: Discussionmentioning

confidence: 99%

Patterning Bacterial Communities on Epithelial Cells

et al. 2013

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Micropatterning of bacteria using aqueous two phase system (ATPS) enables the localized culture and formation of physically separated bacterial communities on human epithelial cell sheets. This method was used to compare the effects of Escherichia coli strain MG1655 and an isogenic invasive counterpart that expresses the invasin (inv) gene from Yersinia pseudotuberculosis on the underlying epithelial cell layer. Large portions of the cell layer beneath the invasive strain were killed or detached while the non-invasive E. coli had no apparent effect on the epithelial cell layer over a 24 h observation period. In addition, simultaneous testing of the localized effects of three different bacterial species; E. coli MG1655, Shigella boydii KACC 10792 and Pseudomonas sp DSM 50906 on an epithelial cell layer is also demonstrated. The paper further shows the ability to use a bacterial predator, Bdellovibrio bacteriovorus HD 100, to selectively remove the E. coli, S. boydii and P. sp communities from this bacteria-patterned epithelial cell layer. Importantly, predation and removal of the P. Sp was critical for maintaining viability of the underlying epithelial cells. Although this paper focuses on a few specific cell types, the technique should be broadly applicable to understand a variety of bacteria-epithelial cell interactions.

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Section: Discussionmentioning

confidence: 99%

Patterning Bacterial Communities on Epithelial Cells

et al. 2013

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“…ATPS‐based cell delivery allows cells to be dispensed into a range of configurations in vitro, including cell colonies, exclusion zones, co‐cultures, heterocellular microarrays, and cells patterned on pre‐existing monolayers (Fang et al, ; Frampton et al, ; Tavana et al, ; Tavana et al, ). However, few studies have examined the ability of this approach to augment growth of cells through confinement in configurations that promote homotypic and heterotypic interactions necessary for appropriate cell growth, differentiation, and function (Joshi, Fuller, Li, & Tavana, ; Joshi, Thakuri, Buchanan, Li, & Tavana, ).…”

Section: Resultsmentioning

confidence: 99%

Precision cell delivery in biphasic polymer systems enhances growth of keratinocytes in culture and promotes their attachment on acellular dermal matrices

Agarwal

Liu

Tam

et al. 2019

J Tissue Eng Regen Med

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Current approaches for precision deposition of cells are not optimized for moist environments or for substrates with complex surface topographic features, for example, the surface of dermal matrices and other biomaterials. To overcome these challenges, an approach is presented that utilizes cell confinement in phase‐separating polymer solutions of polyethylene glycol and dextran to precisely deliver keratinocytes in well‐defined colonies. Using this approach, keratinocyte colonies are produced with superior viability, proliferative capacity, and barrier formation compared with the same number of cells dispersedly seeded across substrate surfaces. It is further demonstrated that keratinocytes delivered in colonies to the surface of acellular dermal matrices form an intact epidermal basal layer more rapidly and more completely than cells delivered by conventional dispersed seeding. These findings demonstrate that delivery of keratinocytes in phase‐separating polymer solutions holds potential for enhancing growth of keratinocytes in culture and production of functional skin equivalents.

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“…[6,26,[31][32][33][34][35][36][37][38] An ATPS is a versatile phase-separated system consisting of different hydrophilic polymers, such as poly(ethylene oxide) (PEO) and dextran (DEX). [6,26,[31][32][33][34][35][36][37][38] An ATPS is a versatile phase-separated system consisting of different hydrophilic polymers, such as poly(ethylene oxide) (PEO) and dextran (DEX).…”

Section: Doi: 101002/adma201904631mentioning

confidence: 99%

Freeform, Reconfigurable Embedded Printing of All‐Aqueous 3D Architectures

et al. 2019

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Aqueous microstructures are challenging to create, handle, and preserve since their surfaces tend to shrink into spherical shapes with minimum surface areas. The creation of freeform aqueous architectures will significantly advance the bioprinting of complex tissue‐like constructs, such as arteries, urinary catheters, and tracheae. The generation of complex, freeform, three‐dimensional (3D) all‐liquid architectures using formulated aqueous two‐phase systems (ATPSs) is demonstrated. These all‐liquid microconstructs are formed by printing aqueous bioinks in an immiscible aqueous environment, which functions as a biocompatible support and pregel solution. By exploiting the hydrogen bonding interaction between polymers in ATPS, the printed aqueous‐in‐aqueous reconfigurable 3D architectures can be stabilized for weeks by the noncovalent membrane at the interface. Different cells can be separately combined with compartmentalized bioinks and matrices to obtain tailor‐designed microconstructs with perfusable vascular networks. The freeform, reconfigurable embedded printing of all‐liquid architectures by ATPSs offers unique opportunities and powerful tools since limitless formulations can be designed from among a breadth of natural and synthetic hydrophilic polymers to mimic tissues. This printing approach may be useful to engineer biomimetic, dynamic tissue‐like constructs for potential applications in drug screening, in vitro tissue models, and regenerative medicine.

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Cell Co-culture Patterning Using Aqueous Two-phase Systems

Cited by 28 publications

References 29 publications

Patterning Bacterial Communities on Epithelial Cells

Patterning Bacterial Communities on Epithelial Cells

Precision cell delivery in biphasic polymer systems enhances growth of keratinocytes in culture and promotes their attachment on acellular dermal matrices

Freeform, Reconfigurable Embedded Printing of All‐Aqueous 3D Architectures

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