Bioreactors for Advanced Cell Culture, (Nano)toxicity, Regenerative Medicine and Organ Bioengineering

Topic Information

Dear Colleagues,

From the micro to the macroscale, bioreactors are widely used for advanced cell cultures. They can be used as tools for regenerative medicine and for generating physiologically relevant in vitro models. Thus, they represent valid animal model alternatives in drug testing, toxicology applications and more generally to investigate the interaction of different substances, nanomaterials, and pathogens with the human body and immune cells. For these applications, biological interfaces such as lung, intestine, skin, and blood vessels, as well as vital organs (liver, heart, brain and kidney) and structural/functional tissues (bone, cartilage, muscles) are of particular interest. Combining engineering and biology, bioreactors are able to replicate key features of organs and tissues and to provide cells with dynamic stimuli such as flow or mechanical and electrical cues. Current challenges relate to the integration of sensing for real-time monitoring of cell conditions and with the definition of novel actuation strategies to imitate physiological deformation mechanisms. Other fundamental innovations regard the combination of bioreactors and biomaterials to mimic tissue and organ 3D architectures and mechanical properties, which are typically time-dependent (viscoelastic) and may evolve over time according to pathophysiological processes. In this context, the Special Issue Bioreactors for Advanced Cell Culture, (Nano)toxicity, Regenerative Medicine and Organ Bioengineering is collecting innovative research articles and targeted reviews that deal with novel and bold applications and with new solutions for improving bioreactor technology. For instance, applications may be related to different areas of regenerative medicine and to nanomaterial toxicology from a molecular and mechanistic perspective. Innovative solutions may include the use of non (or low)-invasive actuation and sensing strategies based on smart materials or systems or the integration of in silico strategies that are able to improve the relevance of the experimental conditions or result in translation.

Dr. Ludovica Cacopardo
Dr. Sandeep Keshavan
Dr. Bharath Babu Nunna
Topic Editors

Keywords

Participating Journals

Journal Name	Impact Factor	CiteScore	Launched Year	First Decision (median)	APC
Applied Sciences applsci	2.7	4.5	2011	16.9 Days	CHF 2400
Bioengineering bioengineering	4.6	4.2	2014	17.7 Days	CHF 2700
Cells cells	6.0	9.0	2012	16.6 Days	CHF 2700
Micromachines micromachines	3.4	4.7	2010	16.1 Days	CHF 2600

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Published Papers (11 papers)

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23 pages, 1050 KiB  

Open AccessReview

Advantages and Potential Benefits of Using Organoids in Nanotoxicology

by Varvara G. Nikonorova, Vladimir V. Chrishtop, Vladimir A. Mironov and Artur Y. Prilepskii

Cells 2023, 12(4), 610; https://0-doi-org.brum.beds.ac.uk/10.3390/cells12040610 - 13 Feb 2023

Cited by 5 | Viewed by 2514

Abstract

Organoids are microtissues that recapitulate the complex structural organization and functions of tissues and organs. Nanoparticles have several specific properties that must be considered when replacing animal models with in vitro studies, such as the formation of a protein corona, accumulation, ability to [...] Read more.

Organoids are microtissues that recapitulate the complex structural organization and functions of tissues and organs. Nanoparticles have several specific properties that must be considered when replacing animal models with in vitro studies, such as the formation of a protein corona, accumulation, ability to overcome tissue barriers, and different severities of toxic effects in different cell types. An increase in the number of articles on toxicology research using organoid models is related to an increase in publications on organoids in general but is not related to toxicology-based publications. We demonstrate how the quantitative assessment of toxic changes in the structure of organoids and the state of their cell collections provide more valuable results for toxicological research and provide examples of research methods. The impact of the tested materials on organoids and their differences are also discussed. In conclusion, we highlight the main challenges, the solution of which will allow researchers to approach the replacement of in vivo research with in vitro research: biobanking and standardization of the structural characterization of organoids, and the development of effective screening imaging techniques for 3D organoid cell organization. Full article

(This article belongs to the Topic Bioreactors for Advanced Cell Culture, (Nano)toxicity, Regenerative Medicine and Organ Bioengineering)

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17 pages, 1155 KiB  

Open AccessReview

Application of Precision-Cut Lung Slices as an In Vitro Model for Research of Inflammatory Respiratory Diseases

by Yan Liu, Ping Wu, Yin Wang, Yansong Liu, Hongfang Yang, Guohua Zhou, Xiaoqi Wu and Qingping Wen

Bioengineering 2022, 9(12), 767; https://0-doi-org.brum.beds.ac.uk/10.3390/bioengineering9120767 - 04 Dec 2022

Cited by 6 | Viewed by 4821

Abstract

The leading cause of many respiratory diseases is an ongoing and progressive inflammatory response. Traditionally, inflammatory lung diseases were studied primarily through animal models, cell cultures, and organoids. These technologies have certain limitations, despite their great contributions to the study of respiratory diseases. [...] Read more.

The leading cause of many respiratory diseases is an ongoing and progressive inflammatory response. Traditionally, inflammatory lung diseases were studied primarily through animal models, cell cultures, and organoids. These technologies have certain limitations, despite their great contributions to the study of respiratory diseases. Precision-cut lung slices (PCLS) are thin, uniform tissue slices made from human or animal lung tissue and are widely used extensively both nationally and internationally as an in vitro organotypic model. Human lung slices bridge the gap between in vivo and in vitro models, and they can replicate the living lung environment well while preserving the lungs’ basic structures, such as their primitive cells and trachea. However, there is no perfect model that can completely replace the structure of the human lung, and there is still a long way to go in the research of lung slice technology. This review details and analyzes the strengths and weaknesses of precision lung slices as an in vitro model for exploring respiratory diseases associated with inflammation, as well as recent advances in this field. Full article

(This article belongs to the Topic Bioreactors for Advanced Cell Culture, (Nano)toxicity, Regenerative Medicine and Organ Bioengineering)

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19 pages, 5338 KiB  

Open AccessArticle

Investigating and Modelling an Engineered Millifluidic In Vitro Oocyte Maturation System Reproducing the Physiological Ovary Environment in the Sheep Model

by Antonella Mastrorocco, Ludovica Cacopardo, Letizia Temerario, Nicola Antonio Martino, Federico Tridente, Annalisa Rizzo, Giovanni Michele Lacalandra, Domenico Robbe, Augusto Carluccio and Maria Elena Dell’Aquila

Cells 2022, 11(22), 3611; https://0-doi-org.brum.beds.ac.uk/10.3390/cells11223611 - 15 Nov 2022

Cited by 3 | Viewed by 1622

Abstract

In conventional assisted reproductive technologies (ARTs), oocytes are in vitro cultured in static conditions. Instead, dynamic systems could better mimic the physiological in vivo environment. In this study, a millifluidic in vitro oocyte maturation (mIVM) system, in a transparent bioreactor integrated with 3D [...] Read more.

In conventional assisted reproductive technologies (ARTs), oocytes are in vitro cultured in static conditions. Instead, dynamic systems could better mimic the physiological in vivo environment. In this study, a millifluidic in vitro oocyte maturation (mIVM) system, in a transparent bioreactor integrated with 3D printed supports, was investigated and modeled thanks to computational fluid dynamic (CFD) and oxygen convection-reaction-diffusion (CRD) models. Cumulus-oocyte complexes (COCs) from slaughtered lambs were cultured for 24 h under static (controls) or dynamic IVM in absence (native) or presence of 3D-printed devices with different shapes and assembly modes, with/without alginate filling. Nuclear chromatin configuration, mitochondria distribution patterns, and activity of in vitro matured oocytes were assessed. The native dynamic mIVM significantly reduced the maturation rate compared to the static group (p < 0.001) and metaphase II (MII) oocytes showed impaired mitochondria distribution (p < 0.05) and activity (p < 0.001). When COCs were included in a combination of concave+ring support, particularly with alginate filling, oocyte maturation and mitochondria pattern were preserved, and bioenergetic/oxidative status was improved (p < 0.05) compared to controls. Results were supported by computational models demonstrating that, in mIVM in biocompatible inserts, COCs were protected from shear stresses while ensuring physiological oxygen diffusion replicating the one occurring in vivo from capillaries. Full article

(This article belongs to the Topic Bioreactors for Advanced Cell Culture, (Nano)toxicity, Regenerative Medicine and Organ Bioengineering)

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18 pages, 3838 KiB  

Open AccessArticle

A Transwell-Based Vascularized Model to Investigate the Effect of Interstitial Flow on Vasculogenesis

by Pengwei Deng, Mengqian Zhao, Xu Zhang and Jianhua Qin

Bioengineering 2022, 9(11), 668; https://0-doi-org.brum.beds.ac.uk/10.3390/bioengineering9110668 - 08 Nov 2022

Viewed by 3838

Abstract

Interstitial flow plays a significant role in vascular system development, mainly including angiogenesis and vasculogenesis. However, compared to angiogenesis, the effect of interstitial flow on vasculogenesis is less explored. Current in vitro models for investigating the effect of interstitial flow on vasculogenesis heavily [...] Read more.

Interstitial flow plays a significant role in vascular system development, mainly including angiogenesis and vasculogenesis. However, compared to angiogenesis, the effect of interstitial flow on vasculogenesis is less explored. Current in vitro models for investigating the effect of interstitial flow on vasculogenesis heavily rely on microfluidic chips, which require microfluidic expertise and facilities, and may not be accessible to biological labs. Here, we proposed a facile approach to building perfusable vascular networks through the self-assembly of endothelial cells in a modified transwell format and investigated the effect of interstitial flow on vasculogenesis. We found that the effect of interstitial flow on vasculogenesis was closely related to the existence of VEGF and fibroblasts in the developed model: (1) In the presence of fibroblasts, interstitial flow (within the range of 0.1–0.6 μm/s) facilitated the perfusability of the engineered vasculatures. Additional VEGF in the culture medium further worked synergically with interstitial flow to develop longer, wider, denser, and more perfusable vasculatures than static counterparts; (2) In the absence of fibroblasts, vasculatures underwent severe regression within 7 days under static conditions. However, interstitial flow greatly inhibited vessel regression and enhanced vascular perfusability and morphogenesis without the need for additional VEGF. These results revealed that the effect of interstitial flow might vary depending on the existence of VEGF and fibroblasts, and would provide some guidelines for constructing in vitro self-assembled vasculatures. The established transwell-based vascularized model provides a simple method to build perfusable vasculatures and could also be utilized for creating functional tissues in regenerative medicine. Full article

(This article belongs to the Topic Bioreactors for Advanced Cell Culture, (Nano)toxicity, Regenerative Medicine and Organ Bioengineering)

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23 pages, 16385 KiB  

Open AccessArticle

Characterization of the Aeration and Hydrodynamics in Vertical-Wheel^™ Bioreactors

by Pedro M. Neto, Diogo E. S. Nogueira, Yas Hashimura, Sunghoon Jung, Bruno Pedras, Mário N. Berberan-Santos, Tiago Palmeira, Brian Lee, Joaquim M. S. Cabral, Vitor Geraldes and Carlos A. V. Rodrigues

Bioengineering 2022, 9(8), 386; https://0-doi-org.brum.beds.ac.uk/10.3390/bioengineering9080386 - 12 Aug 2022

Cited by 3 | Viewed by 2323

Abstract

In this work, the oxygen transport and hydrodynamic flow of the PBS Vertical-Wheel MINI^™ 0.1 bioreactor were characterized using experimental data and computational fluid dynamics simulations. Data acquired from spectroscopy-based oxygenation measurements was compared with data obtained from 3D simulations with a [...] Read more.

In this work, the oxygen transport and hydrodynamic flow of the PBS Vertical-Wheel MINI^™ 0.1 bioreactor were characterized using experimental data and computational fluid dynamics simulations. Data acquired from spectroscopy-based oxygenation measurements was compared with data obtained from 3D simulations with a rigid-lid approximation and LES-WALE turbulence modeling, using the open-source software OpenFOAM-8. The mass transfer coefficients were determined for a range of stirring speeds between 10 and 100 rpm and for working volumes between 60 and 100 mL. Additionally, boundary condition, mesh refinement, and temperature variation studies were performed. Lastly, cell size, energy dissipation rate, and shear stress fields were calculated to determine optimal hydrodynamic conditions for culture. The experimental results demonstrate that the $k_L$ can be predicted using $Sh=1.68R{e}^{0.551}S{c}^{\frac{1}{3}}{G}^{1.18}$, with a mean absolute error of 2.08%. Using the simulations and a correction factor of 0.473, the expression can be correlated to provide equally valid results. To directly obtain them from simulations, a partial slip boundary condition can be tuned, ensuring better near-surface velocity profiles or, alternatively, by deeply refining the mesh. Temperature variation studies support the use of this correlation for temperatures up to 37 °C by using a Schmidt exponent of 1/3. Finally, the flow was characterized as transitional with diverse mixing mechanisms that ensure homogeneity and suspension quality, and the results obtained are in agreement with previous studies that employed RANS models. Overall, this work provides new data regarding oxygen mass transfer and hydrodynamics in the Vertical-Wheel bioreactor, as well as new insights for air-water mass transfer modeling in systems with low interface deformation, and a computational model that can be used for further studies. Full article

(This article belongs to the Topic Bioreactors for Advanced Cell Culture, (Nano)toxicity, Regenerative Medicine and Organ Bioengineering)

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21 pages, 728 KiB  

Open AccessReview

Metabolic Profiling of CHO Cells during the Production of Biotherapeutics

by Mathilde Coulet, Oliver Kepp, Guido Kroemer and Stéphane Basmaciogullari

Cells 2022, 11(12), 1929; https://0-doi-org.brum.beds.ac.uk/10.3390/cells11121929 - 15 Jun 2022

Cited by 19 | Viewed by 8210

Abstract

As indicated by an ever-increasing number of FDA approvals, biotherapeutics constitute powerful tools for the treatment of various diseases, with monoclonal antibodies (mAbs) accounting for more than 50% of newly approved drugs between 2014 and 2018 (Walsh, 2018). The pharmaceutical industry has made [...] Read more.

As indicated by an ever-increasing number of FDA approvals, biotherapeutics constitute powerful tools for the treatment of various diseases, with monoclonal antibodies (mAbs) accounting for more than 50% of newly approved drugs between 2014 and 2018 (Walsh, 2018). The pharmaceutical industry has made great progress in developing reliable and efficient bioproduction processes to meet the demand for recombinant mAbs. Mammalian cell lines are preferred for the production of functional, complex recombinant proteins including mAbs, with Chinese hamster ovary (CHO) cells being used in most instances. Despite significant advances in cell growth control for biologics manufacturing, cellular responses to environmental changes need to be understood in order to further improve productivity. Metabolomics offers a promising approach for developing suitable strategies to unlock the full potential of cellular production. This review summarizes key findings on catabolism and anabolism for each phase of cell growth (exponential growth, the stationary phase and decline) with a focus on the principal metabolic pathways (glycolysis, the pentose phosphate pathway and the tricarboxylic acid cycle) and the families of biomolecules that impact these circuities (nucleotides, amino acids, lipids and energy-rich metabolites). Full article

(This article belongs to the Topic Bioreactors for Advanced Cell Culture, (Nano)toxicity, Regenerative Medicine and Organ Bioengineering)

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25 pages, 6295 KiB  

Open AccessArticle

3D Bioprinting of Prevascularized Full-Thickness Gelatin-Alginate Structures with Embedded Co-Cultures

by Bastian Böttcher, Astrid Pflieger, Jan Schumacher, Berit Jungnickel and Karl-Heinz Feller

Bioengineering 2022, 9(6), 242; https://0-doi-org.brum.beds.ac.uk/10.3390/bioengineering9060242 - 31 May 2022

Cited by 6 | Viewed by 2956

Abstract

The use of bioprinting allows the creation of complex three-dimensional cell laden grafts with spatial placements of different cell lines. However, a major challenge is insufficient nutrient transfer, especially with the increased size of the graft causing necrosis and reduced proliferation. A possibility [...] Read more.

The use of bioprinting allows the creation of complex three-dimensional cell laden grafts with spatial placements of different cell lines. However, a major challenge is insufficient nutrient transfer, especially with the increased size of the graft causing necrosis and reduced proliferation. A possibility to improve nutrient support is the integration of tubular structures for reducing diffusion paths. In this study the influence of prevascularization in full-thickness grafts on cell growth with a variation of cultivation style and cellular composition was investigated. To perform this, the rheological properties of the used gelatin-alginate hydrogel as well as possibilities to improve growth conditions in the hydrogel were assessed. Prevascularized grafts were manufactured using a pneumatic extrusion-based bioprinter with a coaxial extrusion tool. The prevascularized grafts were statically and dynamically cultured with a monoculture of HepG2 cells. Additionally, a co-culture of HepG2 cells, fibroblasts and HUVEC-TERT2 was created while HUVEC-TERT2s were concentrically placed around the hollow channels. A static culture of prevascularized grafts showed short-term improvements in cell proliferation compared to avascular grafts, while a perfusion-based culture showed improvements in mid-term cultivation times. The cultivation of the co-culture indicated the formation of vascular structures from the hollow channels toward avascular areas. According to these results, the integration of prevascular structures show beneficial effects for the in vitro cultivation of bioprinted grafts for which its impact can be increased in larger grafts. Full article

(This article belongs to the Topic Bioreactors for Advanced Cell Culture, (Nano)toxicity, Regenerative Medicine and Organ Bioengineering)

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21 pages, 2503 KiB  

Open AccessReview

Bioengineered Wound Healing Skin Models: The Role of Immune Response and Endogenous ECM to Fully Replicate the Dynamic of Scar Tissue Formation In Vitro

by Francesco Urciuolo, Roberta Passariello, Giorgia Imparato, Costantino Casale and Paolo Antonio Netti

Bioengineering 2022, 9(6), 233; https://0-doi-org.brum.beds.ac.uk/10.3390/bioengineering9060233 - 27 May 2022

Cited by 7 | Viewed by 5384

Abstract

The healing of deep skin wounds is a complex phenomenon evolving according with a fine spatiotemporal regulation of different biological events (hemostasis, inflammation, proliferation, remodeling). Due to the spontaneous evolution of damaged human dermis toward a fibrotic scar, the treatment of deep wounds [...] Read more.

The healing of deep skin wounds is a complex phenomenon evolving according with a fine spatiotemporal regulation of different biological events (hemostasis, inflammation, proliferation, remodeling). Due to the spontaneous evolution of damaged human dermis toward a fibrotic scar, the treatment of deep wounds still represents a clinical concern. Bioengineered full-thickness skin models may play a crucial role in this direction by providing a deep understanding of the process that leads to the formation of fibrotic scars. This will allow (i) to identify new drugs and targets/biomarkers, (ii) to test new therapeutic approaches, and (iii) to develop more accurate in silico models, with the final aim to guide the closure process toward a scar-free closure and, in a more general sense, (iv) to understand the mechanisms involved in the intrinsic and extrinsic aging of the skin. In this work, the complex dynamic of events underlaying the closure of deep skin wound is presented and the engineered models that aim at replicating such complex phenomenon are reviewed. Despite the complexity of the cellular and extracellular events occurring during the skin wound healing the gold standard assay used to replicate such a process is still represented by planar in vitro models that have been largely used to identify the key factors regulating the involved cellular processes. However, the lack of the main constituents of the extracellular matrix (ECM) makes these over-simplistic 2D models unable to predict the complexity of the closure process. Three-dimensional bioengineered models, which aim at recreating the closure dynamics of the human dermis by using exogenous biomaterials, have been developed to fill such a gap. Although interesting mechanistic effects have been figured out, the effect of the inflammatory response on the ECM remodelling is not replicated yet. We discuss how more faithful wound healing models can be obtained by creating immunocompetent 3D dermis models featuring an endogenous ECM. Full article

(This article belongs to the Topic Bioreactors for Advanced Cell Culture, (Nano)toxicity, Regenerative Medicine and Organ Bioengineering)

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18 pages, 3838 KiB  

Open AccessArticle

Preparation of Spheroids from Primary Pig Cells in a Mid-Scale Bioreactor Retaining Their Myogenic Potential

by Katja Stange, Amir Keric, Andreas Friese and Monika Röntgen

Cells 2022, 11(9), 1453; https://0-doi-org.brum.beds.ac.uk/10.3390/cells11091453 - 25 Apr 2022

Cited by 3 | Viewed by 3713

Abstract

Three-dimensional cell culture techniques mimic the in vivo cell environment more adequately than flat surfaces. Spheroids are multicellular aggregates and we aimed to produce scaffold-free spheroids of myogenic origin, called myospheres, using a mid-scale incubator and bioreactor hybrid. For the first time, we [...] Read more.

Three-dimensional cell culture techniques mimic the in vivo cell environment more adequately than flat surfaces. Spheroids are multicellular aggregates and we aimed to produce scaffold-free spheroids of myogenic origin, called myospheres, using a mid-scale incubator and bioreactor hybrid. For the first time, we obtained spheroids from primary porcine muscle cells (PMCs) with this technology and compared their morphology and growth parameters, marker expression, and myogenic potential to C2C12-derived spheroids. Both cell types were able to form round-shaped spheroids in the bioreactor already after 24 h. The mean diameter of the C2C12 spheroids (44.6 µm) was larger than that of the PMCs (32.7 µm), and the maximum diameter exceeded 1 mm. C2C12 cells formed less aggregates than PMCs with a higher packing density (cell nuclei/mm²). After dissociation from the spheroids, C2C12 cells and PMCs started to proliferate again and were able to differentiate into the myogenic lineage, as shown by myotube formation and the expression of F-Actin, Desmin, MyoG, and Myosin. For C2C12, multinucleated syncytia and Myosin expression were observed in spheroids, pointing to accelerated myogenic differentiation. In conclusion, the mid-scale incubator and bioreactor system is suitable for spheroid formation and cultivation from primary muscle cells while preserving their myogenic potential. Full article

(This article belongs to the Topic Bioreactors for Advanced Cell Culture, (Nano)toxicity, Regenerative Medicine and Organ Bioengineering)

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16 pages, 1706 KiB  

Open AccessReview

Novel Techniques and Future Perspective for Investigating Critical-Size Bone Defects

by Elijah Ejun Huang, Ning Zhang, Huaishuang Shen, Xueping Li, Masahiro Maruyama, Takeshi Utsunomiya, Qi Gao, Roberto A. Guzman and Stuart B. Goodman

Bioengineering 2022, 9(4), 171; https://0-doi-org.brum.beds.ac.uk/10.3390/bioengineering9040171 - 11 Apr 2022

Cited by 12 | Viewed by 3792

Abstract

A critical-size bone defect is a challenging clinical problem in which a gap between bone ends will not heal and will become a nonunion. The current treatment is to harvest and transplant an autologous bone graft to facilitate bone bridging. To develop less [...] Read more.

A critical-size bone defect is a challenging clinical problem in which a gap between bone ends will not heal and will become a nonunion. The current treatment is to harvest and transplant an autologous bone graft to facilitate bone bridging. To develop less invasive but equally effective treatment options, one needs to first have a comprehensive understanding of the bone healing process. Therefore, it is imperative to leverage the most advanced technologies to elucidate the fundamental concepts of the bone healing process and develop innovative therapeutic strategies to bridge the nonunion gap. In this review, we first discuss the current animal models to study critical-size bone defects. Then, we focus on four novel analytic techniques and discuss their strengths and limitations. These four technologies are mass cytometry (CyTOF) for enhanced cellular analysis, imaging mass cytometry (IMC) for enhanced tissue special imaging, single-cell RNA sequencing (scRNA-seq) for detailed transcriptome analysis, and Luminex assays for comprehensive protein secretome analysis. With this new understanding of the healing of critical-size bone defects, novel methods of diagnosis and treatment will emerge. Full article

(This article belongs to the Topic Bioreactors for Advanced Cell Culture, (Nano)toxicity, Regenerative Medicine and Organ Bioengineering)

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13 pages, 1790 KiB  

Open AccessArticle

Angiogenesis and Functional Vessel Formation Induced by Interstitial Flow and Vascular Endothelial Growth Factor Using a Microfluidic Chip

by Yufang Liu, Jiao Li, Jiasheng Zhou, Xue Liu, Huibing Li, Yao Lu, Bingcheng Lin, Xiaojie Li and Tingjiao Liu

Micromachines 2022, 13(2), 225; https://0-doi-org.brum.beds.ac.uk/10.3390/mi13020225 - 29 Jan 2022

Cited by 11 | Viewed by 4497

Abstract

Angiogenesis occurs during both physiological and pathological processes. In this study, a microfluidic chip for the development of angiogenesis was utilized to assess angiogenic sprouting and functional vessel formation. We also found that vascular endothelial growth factor (VEGF) was a determinant of the [...] Read more.

Angiogenesis occurs during both physiological and pathological processes. In this study, a microfluidic chip for the development of angiogenesis was utilized to assess angiogenic sprouting and functional vessel formation. We also found that vascular endothelial growth factor (VEGF) was a determinant of the initiation of vascular sprouts, while the direction of these sprouts was greatly influenced by interstitial flow. Isoforms of VEGF such as VEGF₁₂₁, VEGF₁₆₅, and VEGF₁₈₉ displayed different angiogenic properties on the chip as assessed by sprout length and number, vessel perfusion, and connectivity. VEGF₁₆₅ had the highest capacity to induce vascular sprouting among the three isoforms assessed and furthermore, also induced functional vessel formation. This chip could be used to analyze the effect of different angiogenic factors and drugs, as well as to explore the mechanism of angiogenesis induced by such factors. Full article

(This article belongs to the Topic Bioreactors for Advanced Cell Culture, (Nano)toxicity, Regenerative Medicine and Organ Bioengineering)

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