microfluidic perfusion
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2020 ◽  
Author(s):  
Mikhail Nikolaev ◽  
Olga Mitrofanova ◽  
Nicolas Broguiere ◽  
Sara Geraldo ◽  
Yoji Tabata ◽  
...  

Abstract Organoids derived from epithelial stem cells have emerged as powerful platforms to model development and disease in a dish1-3. However, the current mismatch in anatomy, lifespan and size between native organs and their in vitro counterparts severely limits their applicability4. In particular, the closed, cystic architecture of most epithelial stem cell-derived organoids makes experimental manipulation and assay development cumbersome. Here we describe how tissue engineering and cellular self-organization can be combined to guide in vitro organogenesis into openly accessible, functional intestinal tubes termed ‘mini-guts’. Intestinal stem cells (ISCs) rapidly generate simple columnar epithelia when propagated inside basal lamina-like hydrogel scaffolds that feature a tubular and crypt-containing, in vivo-like anatomical structure. Using a microfluidic perfusion system, dead cells shed into the lumen can be continuously removed from the mini-guts. This increases tissue lifespan to months, establishing a homeostatic organoid culture system in which cell proliferation (in crypts) is balanced with cell death (in villus-like domains). The approach developed here can be extended to generate functional tissue/organ models from other epithelial cell types, including primary human stem/progenitor cells from the small intestine, colon or airway, permitting reconstitution of complex organ-level physiology and disease in a personalized manner.



2020 ◽  
Vol 7 (7) ◽  
pp. 200027
Author(s):  
Shinya Yamahira ◽  
Taku Satoh ◽  
Fumiki Yanagawa ◽  
Masato Tamura ◽  
Toshiyuki Takagi ◽  
...  

Inside living organisms, concentration gradients dynamically change over time as biological processes progress. Therefore, methods to construct dynamic microscale concentration gradients in a spatially controlled manner are needed to provide more realistic research environments. Here, we report a novel method for the construction of dynamic microscale concentration gradients in a stepwise manner around cells in micropatterned hydrogel. In our method, cells are encapsulated in a photodegradable hydrogel formed inside a microfluidic perfusion culture device, and perfusion microchannels are then fabricated in the hydrogel by micropatterned photodegradation. The cells in the micropatterned hydrogel can then be cultured by perfusing culture medium through the fabricated microchannels. By using this method, we demonstrate the simultaneous construction of two dynamic concentration gradients, which allowed us to expose the cells encapsulated in the hydrogel to a dynamic microenvironment.



2020 ◽  
Vol 22 (2) ◽  
Author(s):  
Yadir A. Guerrero ◽  
Diti Desai ◽  
Connor Sullivan ◽  
Erick Kindt ◽  
Mary E. Spilker ◽  
...  


Micromachines ◽  
2020 ◽  
Vol 11 (2) ◽  
pp. 150 ◽  
Author(s):  
Baydoun ◽  
Treizeibré ◽  
Follet ◽  
Vanneste ◽  
Creusy ◽  
...  

Ex vivo explant culture models offer unique properties to study complex mechanisms underlying tissue growth, renewal, and disease. A major weakness is the short viability depending on the biopsy origin and preparation protocol. We describe an interphase microfluidic culture system to cultivate full thickness murine colon explants which keeps morphological structures of the tissue up to 192 h. The system was composed of a central well on top of a porous membrane supported by a microchannel structure. The microfluidic perfusion allowed bathing the serosal side while preventing immersion of the villi. After eight days, up to 33% of the samples displayed no histological abnormalities. Numerical simulation of the transport of oxygen and glucose provided technical solutions to improve the functionality of the microdevice.



2020 ◽  
Vol 11 ◽  
pp. 204173142095471 ◽  
Author(s):  
Carlotta Peticone ◽  
David De Silva Thompson ◽  
Nikolay Dimov ◽  
Ben Jevans ◽  
Nick Glass ◽  
...  

Using microspherical scaffolds as building blocks to repair bone defects of specific size and shape has been proposed as a tissue engineering strategy. Here, phosphate glass (PG) microcarriers doped with 5 mol % TiO2 and either 0 mol % CoO (CoO 0%) or 2 mol % CoO (CoO 2%) were investigated for their ability to support osteogenic and vascular responses of human mesenchymal stem cells (hMSCs). Together with standard culture techniques, cell-material interactions were studied using a novel perfusion microfluidic bioreactor that enabled cell culture on microspheres, along with automated processing and screening of culture variables. While titanium doping was found to support hMSCs expansion and differentiation, as well as endothelial cell-derived vessel formation, additional doping with cobalt did not improve the functionality of the microspheres. Furthermore, the microfluidic bioreactor enabled screening of culture parameters for cell culture on microspheres that could be potentially translated to a scaled-up system for tissue-engineered bone manufacturing.



The Analyst ◽  
2020 ◽  
Vol 145 (14) ◽  
pp. 4815-4826 ◽  
Author(s):  
Emily L. Jackson-Holmes ◽  
Amanda W. Schaefer ◽  
Todd C. McDevitt ◽  
Hang Lu

This work explores how media exchange frequency and device geometry modulate the biochemical environment and impact three-dimensional stem cell differentiation.



2019 ◽  
Vol 30 (1) ◽  
pp. 1908349 ◽  
Author(s):  
Liliang Ouyang ◽  
James P. K. Armstrong ◽  
Qu Chen ◽  
Yiyang Lin ◽  
Molly M. Stevens


2019 ◽  
Vol 26 (1) ◽  
Author(s):  
Jyong-Huei Lee ◽  
Kuan-Lun Ho ◽  
Shih-Kang Fan

Abstract Engineering approaches were adopted for liver microsystems to recapitulate cell arrangements and culture microenvironments in vivo for sensitive, high-throughput and biomimetic drug screening. This review introduces liver microsystems in vitro for drug hepatotoxicity, drug-drug interactions, metabolic function and enzyme induction, based on cell micropatterning, hydrogel biofabrication and microfluidic perfusion. The engineered microsystems provide varied microenvironments for cell culture that feature cell coculture with non-parenchymal cells, in a heterogeneous extracellular matrix and under controllable perfusion. The engineering methods described include cell micropatterning with soft lithography and dielectrophoresis, hydrogel biofabrication with photolithography, micromolding and 3D bioprinting, and microfluidic perfusion with endothelial-like structures and gradient generators. We discuss the major challenges and trends of liver microsystems to study drug response in vitro.





2019 ◽  
Author(s):  
Julia Rogal ◽  
Carina Binder ◽  
Elena Kromidas ◽  
Christopher Probst ◽  
Stefan Schneider ◽  
...  

AbstractObesity and its numerous adverse health consequences have taken on global, pandemic proportions. White adipose tissue (WAT) – a key contributor in many metabolic diseases – contributes about one fourth of a healthy human’s body mass. Despite its significance, many WAT-related pathophysiogical mechanisms in humans are still not understood, largely due to the reliance on non-human animal models. In recent years, Organ-on-a-chip (OoC) platforms have developed into promising alternatives for animal models; these systems integrate engineered human tissues into physiological microenvironment supplied by a vasculature-like microfluidic perfusion. Here, we report the development of a novel OoC that integrates functional mature human WAT. The WAT-on-a-chip is a multilayer device that features tissue chambers tailored specifically for the maintenance of 3D tissues based on human primary adipocytes, with supporting nourishment provided through perfused media channels. The platform’s capability to maintain long-term viability and functionality of WAT was confirmed by real-time monitoring of fatty acid uptake, by quantification of metabolite release into the effluent media as well as by an intact responsiveness to a therapeutic compound. The novel system provides a promising tool for wide-ranging applications in mechanistic research of WAT-related biology, in studying of pathophysiological mechanisms in obesity and diabetes, and in R&D of pharmaceutical industry.



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