scholarly journals In vivo–mimicking microfluidic perfusion culture of pancreatic islet spheroids

2019 ◽  
Vol 5 (11) ◽  
pp. eaax4520 ◽  
Author(s):  
Yesl Jun ◽  
JaeSeo Lee ◽  
Seongkyun Choi ◽  
Ji Hun Yang ◽  
Maike Sander ◽  
...  
Keyword(s):  
Drug Testing ◽  
Cell Damage ◽  
Three Dimensional ◽  
Perfusion Culture ◽  
Interstitial Flow ◽  
Static Culture ◽  
Culture Models ◽  
Main Component

Native pancreatic islets interact with neighboring cells by establishing three-dimensional (3D) structures, and are surrounded by perfusion at an interstitial flow level. However, flow effects are generally ignored in islet culture models, although cell perfusion is known to improve the cell microenvironment and to mimic in vivo physiology better than static culture systems. Here, we have developed functional islet spheroids using a microfluidic chip that mimics interstitial flow conditions with reduced shear cell damage. Dynamic culture, compared to static culture, enhanced islet health and maintenance of islet endothelial cells, reconstituting the main component of islet extracellular matrix within spheroids. Optimized flow condition allowed localization of secreted soluble factors near spheroids, facilitating diffusion-mediated paracrine interactions within islets, and enabled long-term maintenance of islet morphology and function for a month. The proposed model can aid islet preconditioning before transplantation and has potential applications as an in vitro model for diabetic drug testing.

scholarly journals Bonding of Flexible Membranes for Perfusable Vascularized Networks Patch

2021 ◽  
Author(s):  
Soyoung Hong ◽  
Yejin Song ◽  
Jaesoon Choi ◽  
Changmo Hwang
Keyword(s):  
Endothelial Cells ◽  
Three Dimensional ◽  
Perfusion Culture ◽  
Vascular Network ◽  
Electrospun Membrane ◽  
Static Culture ◽  
Flexible Membranes ◽  
Stress Related Genes

Abstract BACKGROUND: In vitro generation of three-dimensional vessel network is crucial to investigate and possibly improve vascularization after implantation in vivo. This work has the purpose of engineering complex tissue regeneration of a vascular network including multiple cell-type, an extracellular matrix, and perfusability for clinical application. METHODS: The two electrospun membranes bonded with the vascular network shape are cultured with endothelial cells and medium flow through the engineered vascular network. The flexible membranes are bonded by amine-epoxy reaction and examined the perfusability with fluorescent beads. Also, the perfusion culture for 7 days of the endothelial cells is compared with static culture on the engineered vascular network membrane. RESULTS: The engineered membranes are showed perfusability through the vascular network, and the perfused network resulted in more cell proliferation and variation of the shear stress-related genes expression compared to the static culture. Also, for the generation of the complex vascularized network, pericytes are co-cultured with the engineered vascular network, which results in the Collagen I is expressed on the outer surface of the engineered structure. CONCLUSION: This study is showing the perfusable in vitro engineered vascular network with electrospun membrane. In further, the 3D vascularized network module can be expected as a platform for drug screening and regenerative medicine.

Comparison of two-dimensional (2D)- and three-dimensional (3D)-culture models as drug testing platforms in GIST: experience with axitinib in vitro

2017 ◽  
Vol 72 ◽  
pp. S155
Author(s):  
V. Martínez-Marín ◽  
A. Redondo ◽  
V. Heredia ◽  
L. Guerra ◽  
M. Miguel-Martín ◽  
...  
Keyword(s):  
Drug Testing ◽  
Three Dimensional ◽  
3D Culture ◽  
Two Dimensional ◽  
Culture Models

scholarly journals Current Advances in 3D Tissue and Organ Reconstruction

2021 ◽  
Vol 22 (2) ◽  
pp. 830
Author(s):  
Georgia Pennarossa ◽  
Sharon Arcuri ◽  
Teresina De Iorio ◽  
Fulvio Gandolfi ◽  
Tiziana A. L. Brevini
Keyword(s):  
Cell Biology ◽  
Drug Testing ◽  
Three Dimensional ◽  
3D Culture ◽  
In Vitro Models ◽  
The Body ◽  
Cellular Functions ◽  
Culture Systems

Bi-dimensional culture systems have represented the most used method to study cell biology outside the body for over a century. Although they convey useful information, such systems may lose tissue-specific architecture, biomechanical effectors, and biochemical cues deriving from the native extracellular matrix, with significant alterations in several cellular functions and processes. Notably, the introduction of three-dimensional (3D) platforms that are able to re-create in vitro the structures of the native tissue, have overcome some of these issues, since they better mimic the in vivo milieu and reduce the gap between the cell culture ambient and the tissue environment. 3D culture systems are currently used in a broad range of studies, from cancer and stem cell biology, to drug testing and discovery. Here, we describe the mechanisms used by cells to perceive and respond to biomechanical cues and the main signaling pathways involved. We provide an overall perspective of the most recent 3D technologies. Given the breadth of the subject, we concentrate on the use of hydrogels, bioreactors, 3D printing and bioprinting, nanofiber-based scaffolds, and preparation of a decellularized bio-matrix. In addition, we report the possibility to combine the use of 3D cultures with functionalized nanoparticles to obtain highly predictive in vitro models for use in the nanomedicine field.

scholarly journals Patient-Derived Organoids as a Model for Cancer Drug Discovery

2021 ◽  
Vol 22 (7) ◽  
pp. 3483
Author(s):  
Colin Rae ◽  
Francesco Amato ◽  
Chiara Braconi
Keyword(s):  
Drug Testing ◽  
Ex Vivo ◽  
Three Dimensional ◽  
Tumour Microenvironment ◽  
In Vitro Models ◽  
Personalized Therapy ◽  
Cancer Drug ◽  
Patient Response

In the search for the ideal model of tumours, the use of three-dimensional in vitro models is advancing rapidly. These are intended to mimic the in vivo properties of the tumours which affect cancer development, progression and drug sensitivity, and take into account cell–cell interactions, adhesion and invasiveness. Importantly, it is hoped that successful recapitulation of the structure and function of the tissue will predict patient response, permitting the development of personalized therapy in a timely manner applicable to the clinic. Furthermore, the use of co-culture systems will allow the role of the tumour microenvironment and tissue–tissue interactions to be taken into account and should lead to more accurate predictions of tumour development and responses to drugs. In this review, the relative merits and limitations of patient-derived organoids will be discussed compared to other in vitro and ex vivo cancer models. We will focus on their use as models for drug testing and personalized therapy and how these may be improved. Developments in technology will also be considered, including the use of microfluidics, 3D bioprinting, cryopreservation and circulating tumour cell-derived organoids. These have the potential to enhance the consistency, accessibility and availability of these models.

scholarly journals Investigating PEGDA and GelMA Microgel Models for Sustained 3D Heterotypic Dermal Papilla and Keratinocyte Co-Cultures

2021 ◽  
Vol 22 (4) ◽  
pp. 2143 ◽  
Author(s):  
Justin J.Y. Tan ◽  
Duc-Viet Nguyen ◽  
John E. Common ◽  
Chunyong Wu ◽  
Paul C.L. Ho ◽  
...  
Keyword(s):  
Hair Follicle ◽  
Three Dimensional ◽  
Dermal Papilla ◽  
Glycol Diacrylate ◽  
Prolonged Duration ◽  
Culture Models ◽  
Poly Ethylene

Hair follicle morphogenesis is heavily dependent on reciprocal, sequential, and epithelial-mesenchymal interaction (EMI) between epidermal stem cells and the specialized cells of the underlying mesenchyme, which aggregate to form the dermal condensate (DC) and will later become the dermal papilla (DP). Similar models were developed with a co-culture of keratinocytes and DP cells. Previous studies have demonstrated that co-culture with keratinocytes maintains the in vivo characteristics of the DP. However, it is often challenging to develop three-dimensional (3D) DP and keratinocyte co-culture models for long term in vitro studies, due to the poor intercellular adherence between keratinocytes. Keratinocytes exhibit exfoliative behavior, and the integrity of the DP and keratinocyte co-cultured spheroids cannot be maintained over prolonged culture. Short durations of culture are unable to sufficiently allow the differentiation and re-programming of the keratinocytes into hair follicular fate by the DP. In this study, we explored a microgel array approach fabricated with two different hydrogel systems. Using poly (ethylene glycol) diacrylate (PEGDA) and gelatin methacrylate (GelMA), we compare their effects on maintaining the integrity of the cultures and their expression of important genes responsible for hair follicle morphogenesis, namely Wnt10A, Wnt10B, and Shh, over prolonged duration. We discovered that low attachment surfaces such as PEGDA result in the exfoliation of keratinocytes and were not suitable for long-term culture. GelMA, on the hand, was able to sustain the integrity of co-cultures and showed higher expression of the morphogens overtime.

scholarly journals Novel Bioengineered Three-Dimensional Human Intestinal Model for Long-Term Infection of Cryptosporidium parvum

2017 ◽  
Vol 85 (3) ◽  
Author(s):  
Maria A DeCicco RePass ◽  
Ying Chen ◽  
Yinan Lin ◽  
Wenda Zhou ◽  
David L. Kaplan ◽  
...  
Keyword(s):  
Cryptosporidium Parvum ◽  
Drug Targets ◽  
Diarrheal Disease ◽  
Three Dimensional ◽  
Content Type ◽  
Tissue Model ◽  
Culture Models

ABSTRACT Cryptosporidium spp. are apicomplexan parasites of global importance that cause human diarrheal disease. In vitro culture models that may be used to study this parasite and that have physiological relevance to in vivo infection remain suboptimal. Thus, the pathogenesis of cryptosporidiosis remains poorly characterized, and interventions for the disease are limited. In this study, we evaluated the potential of a novel bioengineered three-dimensional (3D) human intestinal tissue model (which we developed previously) to support long-term infection by Cryptosporidium parvum. Infection was assessed by immunofluorescence assays and confocal and scanning electron microscopy and quantified by quantitative reverse transcription-PCR. We found that C. parvum infected and developed in this tissue model for at least 17 days, the extent of the study time used in the present study. Contents from infected scaffolds could be transferred to fresh scaffolds to establish new infections for at least three rounds. Asexual and sexual stages and the formation of new oocysts were observed during the course of infection. Additionally, we observed ablation, blunting, or distortion of microvilli in infected epithelial cells. Ultimately, a 3D model system capable of supporting continuous Cryptosporidium infection will be a useful tool for the study of host-parasite interactions, identification of putative drug targets, screening of potential interventions, and propagation of genetically modified parasites.

scholarly journals Microfluidic device to control interstitial flow-mediated homotypic and heterotypic cellular communication

Lab on a Chip ◽  
2015 ◽  
Vol 15 (17) ◽  
pp. 3521-3529 ◽  
Author(s):  
Luis F. Alonzo ◽  
Monica L. Moya ◽  
Venktesh S. Shirure ◽  
Steven C. George
Keyword(s):  
Microfluidic Device ◽  
Three Dimensional ◽  
Cellular Communication ◽  
Interstitial Flow ◽  
Temporal Interactions

To address the gap between in vivo microenvironments and in vitro systems, we have developed a novel microfluidic device that precisely controls the spatial and temporal interactions between adjacent three-dimensional cellular environments.

scholarly journals 3D Bioprinting of Vascularized Tissues for in vitro and in vivo Applications

2021 ◽  
Vol 9 ◽  
Author(s):  
Earnest P. Chen ◽  
Zeren Toksoy ◽  
Bruce A. Davis ◽  
John P. Geibel
Keyword(s):  
Tissue Engineering ◽  
Drug Testing ◽  
Rapid Development ◽  
Three Dimensional ◽  
3D Bioprinting ◽  
Vascular Networks ◽  
Main Challenge ◽  
3D Printed

With a limited supply of organ donors and available organs for transplantation, the aim of tissue engineering with three-dimensional (3D) bioprinting technology is to construct fully functional and viable tissue and organ replacements for various clinical applications. 3D bioprinting allows for the customization of complex tissue architecture with numerous combinations of materials and printing methods to build different tissue types, and eventually fully functional replacement organs. The main challenge of maintaining 3D printed tissue viability is the inclusion of complex vascular networks for nutrient transport and waste disposal. Rapid development and discoveries in recent years have taken huge strides toward perfecting the incorporation of vascular networks in 3D printed tissue and organs. In this review, we will discuss the latest advancements in fabricating vascularized tissue and organs including novel strategies and materials, and their applications. Our discussion will begin with the exploration of printing vasculature, progress through the current statuses of bioprinting tissue/organoids from bone to muscles to organs, and conclude with relevant applications for in vitro models and drug testing. We will also explore and discuss the current limitations of vascularized tissue engineering and some of the promising future directions this technology may bring.

Engineering a Three-Dimensional In Vitro Drug Testing Platform for Glioblastoma

2015 ◽  
Vol 6 (4) ◽  
Author(s):  
Metin Akay ◽  
Duong T. Nguyen ◽  
Yantao Fan ◽  
Yasemin M. Akay
Keyword(s):  
Cell Culture ◽  
Drug Testing ◽  
Three Dimensional ◽  
In Vitro Tests ◽  
Valuable Insight ◽  
Poly Ethylene Glycol ◽  
Poly Ethylene ◽  
Cancer Spheroids

Three-dimensional (3D) in vivo cell culture modeling is quickly emerging as a platform to replace two-dimensional (2D) monolayer cell culture in vitro tests. Three-dimensional tumor models mimic physiological conditions and provide valuable insight of the tumor cell response to drug discovery application. In this study, we used poly(ethylene glycol) (PEG) hydrogel microwells to generate 3D brain cancer spheroids and studied their treatment with anticancer drugs in single or combination treatment. Glioblastoma (GBM) spheroids were grown through 14 days before infecting with two drugs: Pitavastatin and Irinotecan at various concentrations. A significant cell lysis was observed and cell viability decreased to lower than 7% when drugs were combined at the concentration Pitavastatin 10 μM and Irinotecan 50 μM to infect after 7 days. These findings demonstrate a promising platform—PEG hydrogel microwells—that should be an efficient way to test the drug sensitivity in vitro as well as application in different studies.

scholarly journals Organs-on-a-chip

2017 ◽  
Vol 1 ◽  
pp. 239784731772635 ◽  
Author(s):  
David Bovard ◽  
Anita Iskandar ◽  
Karsta Luettich ◽  
Julia Hoeng ◽  
Manuel C Peitsch
Keyword(s):  
Biological Effects ◽  
Three Dimensional ◽  
In Vitro Models ◽  
New Drugs ◽  
New Paradigm ◽  
Mechanisms Of Toxicity ◽  
Culture Models ◽  
Dose Responses

In the last few years, considerable attention has been given to in vitro models in an attempt to reduce the use of animals and to decrease the rate of preclinical failure associated with the development of new drugs. Simple two-dimensional cultures grown in a dish are now frequently replaced by organotypic cultures with three-dimensional (3-D) architecture, which enables interactions between cells, promoting their differentiation and increasing their in vivo likeness. Microengineering now enables the incorporation of small devices into 3-D culture models to reproduce the complex microenvironment of the modeled organ, often referred to as organs-on-a-chip (OoCs). This review describes various OoCs developed to mimic liver, brain, kidney, and lung tissues. Current challenges encountered in attempts to recreate the in vivo environment are described, as well as some examples of OoCs. Finally, attention is given to the ongoing evolution of OoCs with the aim of solving one of the major limitations in that they can only represent a single organ. Multi-organ-on-a-chip (MOC) systems mimic organ interactions observed in the human body and aim to provide the features of compound uptake, metabolism, and excretion, while simultaneously allowing for insights into biological effects. MOCs might therefore represent a new paradigm in drug development, providing a better understanding of dose responses and mechanisms of toxicity, enabling the detection of drug resistance and supporting the evaluation of pharmacokinetic–pharmacodynamics parameters.

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