Microfluidic analysis of heterotypic cellular interactions: A review of techniques and applications

doi:10.1016/j.trac.2019.03.026

TrAC Trends in Analytical Chemistry

Volume 117, August 2019, Pages 166-185

https://doi.org/10.1016/j.trac.2019.03.026 Get rights and content

Highlights

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The importance of heterotypic cellular communications and the need for co-culture systems are highlighted.
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Different microfluidic methods for analyzing heterotypic cell interactions are discussed.
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Recently developed in vitro organ models are classified based on different microfluidic co-culturing methods.
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Microfluidic co-culture models for various diseases including cancer metastasis and drug screening are reviewed.

Abstract

Analyzing heterotypic cellular communications is essential to understand various physiological and pathological mechanisms. Although static, 2-dimensional in vitro cell monoculture models allow a high degree of experimental control, they poorly approximate in vivo conditions due to the lack of heterotypic cellular interactions. Co-culture systems permit such interactions and recently, microfluidic platforms have been significantly used for co-culturing due to their various benefits in closely mimicking in vivo microenvironment. This review paper outlines the factors influencing co-culture systems and identifies the available engineering methods for forming co-cultures. The paper also reviews the different microfluidic co-culturing techniques based on conventional, compartmentalized and droplet microfluidic methods and discuss their applications in engineering organ and disease models and drug screening. The categorization of recently developed in vitro organ and disease models based on the different microfluidic co-culture techniques will serve as a guideline for choosing the appropriate method for an intended cell-cell communication study.

Introduction

Cell culturing is a technique for cultivating cells outside a natural biological system. Cells can proliferate in an environment that resembles in vivo conditions, when necessary biochemical and physicochemical factors are presented. Despite the advances in cell culturing techniques, reliability of the results and their applicability to natural complex biological systems is still debated, and as a result, there are still needs for better methods to simulate in vivo conditions [1]. Cellular communication plays a major role in regulating cellular functions such as homeostasis, regeneration and healing processes [2]. Cells in vivo have a complex organization in the form of two-dimensional (2D) or three-dimensional (3D) microstructures and exist in a dynamic environment with constant intercommunication [3]. These communications can be either between same type of cells (homotypic) or between different types of cells (heterotypic). These interactions are mediated either through direct contact of cells or exchange of soluble factors within their microenvironments. However, cells grown in vitro lack these communications and are also exposed to a static and homogeneous environment which is not representing in vivo conditions [4], [5]. The growth rate, morphology or intracellular metabolic activities of cells in vitro are eventually altered [6]. To closely mimic in vivo conditions and facilitate cellular crosstalk, co-culturing techniques have been emerged.

At the early stage of co-culture, culturing multiple cells on a same platform was difficult, as it increased the complexity of the platform to handle multiple cells at a time. Co-culture studies at the beginning seeded multiple cells randomly on a culturing substrate, which hindered the control over the local cell seeding density and the extent of cell-cell communication [7], [8]. However, the recent advances in the engineering tools and methods have enabled us to handle multiple cells easier, facilitating cell co-culturing [9]. Microfabrication technologies have been developed to fabricate the patterned co-culture platforms providing features comparable to cellular dimensions. Hence, the ability to precisely manipulate the cellular microenvironment and intercellular interaction has been enhanced to configure multiple cells in 2D/2D, 2D/3D or 3D/3D arrangements [10].

Recently, microfluidic platforms built by the microfabrication technology have been used as a promising tool for in vitro cell patterning. In such platforms, soft lithography (which uses elastomeric materials) has been employed to fabricate and replicate micro and nanostructures which could be used for cell patterning. As a result, channels and chambers in a micron scale have been fabricated to ensure controlled and continuous perfusion of media, required for providing a dynamic microenvironment. In addition, these platforms make it possible to mimic complex tissue structures by patterning multiple types of cells in multiple channels. Due to aforementioned benefits, this technology provides cost-effective, integrated, high-throughput cell culture systems [11], [12], [13], [14], [15], [16]. Thus, this review paper focuses on the importance of microfluidics as the tool of choice for analyzing heterotypic cellular interactions through co-culture setups. The review aims to give a comprehensive knowledge of different types of microfluidic co-culturing platforms based on conventional, compartmentalization and droplet microfluidic methods. We also discuss the role of the microfluidic co-culture platforms in recent applications such as organ and disease modeling, cancer-related studies, and drug screening. Hence, the classification of the existing microfluidic co-culture methods for a variety of biological and biomedical applications can help researchers choose an appropriate method for their intended cell-cell communication studies.

Section snippets

Factors for co-culturing cells

Stability of co-culture systems is controlled through the heterotypic communication between target and assisting cell populations. Based on their functionality, a distinct cell population can be classified as either the target cells or assisting cells in a co-culture system [17]. The target cells form the part of the desired tissue and are essential for the tissue to function. There are also studies of multiple target cell types co-cultured together where one assisted the other. The assisting

Microfluidic co-culture techniques to analyze heterotypic cellular interactions

Microfluidic systems have become an effective platform for various applications across multiple fields. In particular, it offers notable advantages for cell culturing. Conventional in vitro cell cultures and the other soft lithographic techniques provide only static cell culture conditions, whereas microfluidic systems can provide dynamic cell culture microenvironments due to the continuous perfusion of gases and nutrients, better mimicking in vivo conditions [12], [92]. Apart from the easy

Disease modeling

Existing organ-on-chip platforms have been utilized for disease modeling. Since organs respond diversely to various diseases that might arise from different locations of the organs, the complexity of the in vivo disease mechanisms makes it difficult to develop therapeutic options. The initial step in building a disease model is to engineer an organ construct. This is followed by forming complex environments mimicking the cause and progression of disease. Besides the organ-specific disease

Concluding remarks and future prospects

Co-culturing multiple types of cells and heterotypic cellular interactions offer structural complexity as well as bio-physicochemical factors that cells require to improve tissue functions. We discussed the variables that are considered to set up a co-culture system. The strategies for micropatterned co-culture were explored, which generally have a better spatiotemporal control in terms of the cellular interaction compared to the random co-culture. It shows that the microfluidic co-culture

Acknowledgement

This work was supported by the Natural Sciences and Engineering Research Council of Canada (NSERC) Discovery Grants and Canada Foundation for Innovation John R. Evans Leaders Opportunity Fund.

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Microfluidic analysis of heterotypic cellular interactions: A review of techniques and applications

Highlights

Abstract

Introduction

Section snippets

Factors for co-culturing cells

Microfluidic co-culture techniques to analyze heterotypic cellular interactions

Disease modeling

Concluding remarks and future prospects

Acknowledgement

J. Pharm. Sci.

Exp. Cell Res.

Anal. Chim. Acta

Adv. Drug Deliv. Rev.

J. Neurosci. Methods

Acta Biomater.

Biomaterials

Biomaterials

Lab. Investig.

J. Invest. Dermatol.

Exp. Cell Res.

J. Biol. Chem.

Biochem. Biophys. Res. Commun.

Stem Cell Res.

Biochim. Biophys. Acta Gen. Subj.

Biochem. Biophys. Res. Commun.

Acta Biomater.

Trends Biotechnol.

Biomaterials

Biomaterials

Biochem. Biophys. Res. Commun.

J. Photochem. Photobiol. C Photochem. Rev.

Biomaterials

Biosens. Bioelectron.

Biosens. Bioelectron.

Acta Biomater.

Curr. Opin. Biotechnol.

Curr. Opin. Genet. Dev.

Adv. Drug Deliv. Rev.

Probing cell–cell communication with microfluidic devices

Lab Chip

Microfluidic devices for studying heterotypic cell-cell interactions and tissue specimen cultures under controlled microenvironments

Biomicrofluidics

Beyond the petri dish

Nat. Biotechnol.

Building drug delivery into tissue engineering design

Nat. Rev. Drug Discov.

Microfluidic cell culture systems for cellular analysis

Biochip J.

Microengineering of cellular interactions

Annu. Rev. Biomed. Eng.

Co-culture systems and technologies: taking synthetic biology to the next level

J. R. Soc. Interface

Microfluidic cell culture systems for cellular analysis

Biochip J.

Cells on chips

Nature

The origins and the future of microfluidics

Nature

Microfluidics: fluid physics at the nanoliter scale

Rev. Mod. Phys.

Micro-Electro-mechanical-systems (MEMS) and fluid flows

Annu. Rev. Fluid Mech.

Microfluidics: basic issues, applications, and challenges

AIChE J.

Advances in tissue engineering through stem cell-based co-culture

J. Tissue Eng. Regen. Med.

Stable coexistence of two Caldicellulosiruptor species in a de novo constructed hydrogen-producing co-culture

Microb. Cell Fact.

Building communities one bacterium at a time

Proc. Natl. Acad. Sci.

Collective motion of cells mediates segregation and pattern formation in co-cultures

PLoS One

Drug metabolizing enzymes in rat hepatocytes co-cultured with cell lines, in vitro cell

Dev. Biol.

Microfabrication in Tissue Engineering and Bioartificial Organs

Liver sinusoidal endothelial cells: isolation, purification, characterization and interaction with hepatocytes

Revis. Sobre Biol. Cell. RBC

Induction of glutamine synthetase in periportal hepatocytes by cocultivation with a liver epithelial cell line

Eur. J. Cell Biol.