bims-orenst 2022-03-13 papers

bims-orenst

Biomed News

on Organs-on-chips and engineered stem cell models

Issue of 2022‒03‒13
eight papers selected by
Joram Mooiweer
University of Groningen

Establishment of physiologically relevant oxygen gradients in microfluidic organ chips.
A neurovascular unit-on-a-chip: culture and differentiation of human neural stem cells in a three-dimensional microfluidic environment.
An In Vitro Microfluidic Alveolus Model to Study Lung Biomechanics.
Alternative Brain Slice-on-a-Chip for Organotypic Culture and Effective Fluorescence Injection Testing.
Brain organoid-on-chip system to study the effects of breast cancer derived exosomes on the neurodevelopment of brain.
Bordetella pertussis-infected innate immune cells drive the anti-pertussis response of human airway epithelium.
Cyclic Stretching Induces Maturation of Human-Induced Pluripotent Stem Cell-Derived Cardiomyocytes through Nuclear-Mechanotransduction.
Organoid screening reveals epigenetic vulnerabilities in human colorectal cancer.

Lab Chip. 2022 Mar 11.

Establishment of physiologically relevant oxygen gradients in microfluidic organ chips.

Jennifer Grant, Elizabeth Lee, Micaela Almeida, Seongmin Kim, Nina LoGrande, Girija Goyal, Adama Marie Sesay, David T Breault, Rachelle Prantil-Baun, Donald E Ingber.

In vitro models of human organs must accurately reconstitute oxygen concentrations and gradients that are observed in vivo to mimic gene expression, metabolism, and host-microbiome interactions. Here we describe a simple strategy to achieve physiologically relevant oxygen tension in a two-channel human small intestine-on-a-chip (Intestine Chip) lined with primary human duodenal epithelium and intestinal microvascular endothelium in parallel channels separated by a porous membrane while both channels are perfused with oxygenated medium. This strategy was developed using computer simulations that predicted lowering the oxygen permeability of poly-dimethylsiloxane (PDMS) chips in specified locations using a gas impermeable film will allow the cells to naturally decrease the oxygen concentration through aerobic respiration and reach steady-state oxygen levels <36 mm Hg (<5%) within the epithelial lumen. The approach was experimentally confirmed using chips with embedded oxygen sensors that maintained this stable oxygen gradient. Furthermore, Intestine Chips cultured with this approach supported formation of a villus epithelium interfaced with a continuous endothelium and maintained intestinal barrier integrity for 72 h. This strategy recapitulates in vivo functionality in an efficient, inexpensive, and scalable format that improves the robustness and translatability of Organ Chip technology for studies on microbiome as well as oxygen sensitivity.

DOI: https://doi.org/10.1039/d2lc00069e
Neural Regen Res. 2022 Oct;17(10): 2260-2266

A neurovascular unit-on-a-chip: culture and differentiation of human neural stem cells in a three-dimensional microfluidic environment.

Wen-Juan Wei, Ya-Chen Wang, Xin Guan, Wei-Gong Chen, Jing Liu.

  Biological studies typically rely on a simple monolayer cell culture, which does not reflect the complex functional characteristics of human tissues and organs, or their real response to external stimuli. Microfluidic technology has advantages of high-throughput screening, accurate control of the fluid velocity, low cell consumption, long-term culture, and high integration. By combining the multipotential differentiation of neural stem cells with high throughput and the integrated characteristics of microfluidic technology, an in vitro model of a functionalized neurovascular unit was established using human neural stem cell-derived neurons, astrocytes, oligodendrocytes, and a functional microvascular barrier. The model comprises a multi-layer vertical neural module and vascular module, both of which were connected with a syringe pump. This provides controllable conditions for cell inoculation and nutrient supply, and simultaneously simulates the process of ischemic/hypoxic injury and the process of inflammatory factors in the circulatory system passing through the blood-brain barrier and then acting on the nerve tissue in the brain. The in vitro functionalized neurovascular unit model will be conducive to central nervous system disease research, drug screening, and new drug development.

Keywords:  (neural) differentiation; astrocyte; blood-brain barrier; brain microvascular endothelial cells; central nervous system; microfluidics; neural stem cells; neuron; neurovascular unit; oligodendrocyte; organ-on-a-chip

DOI:  https://doi.org/10.4103/1673-5374.337050
Front Bioeng Biotechnol. 2022 ;10 848699

An In Vitro Microfluidic Alveolus Model to Study Lung Biomechanics.

Vardhman Kumar, Sajeesh Kumar Madhurakkat Perikamana, Aleksandra Tata, Jiaul Hoque, Anna Gilpin, Purushothama Rao Tata, Shyni Varghese.

  The gas exchange units of the lung, the alveoli, are mechanically active and undergo cyclic deformation during breathing. The epithelial cells that line the alveoli contribute to lung function by reducing surface tension via surfactant secretion, which is highly influenced by the breathing-associated mechanical cues. These spatially heterogeneous mechanical cues have been linked to several physiological and pathophysiological states. Here, we describe the development of a microfluidically assisted lung cell culture model that incorporates heterogeneous cyclic stretching to mimic alveolar respiratory motions. Employing this device, we have examined the effects of respiratory biomechanics (associated with breathing-like movements) and strain heterogeneity on alveolar epithelial cell functions. Furthermore, we have assessed the potential application of this platform to model altered matrix compliance associated with lung pathogenesis and ventilator-induced lung injury. Lung microphysiological platforms incorporating human cells and dynamic biomechanics could serve as an important tool to delineate the role of alveolar micromechanics in physiological and pathological outcomes in the lung.

Keywords:  in vitro system; lung; microfluidics; microphysiological system; organ-on-a-chip

DOI:  https://doi.org/10.3389/fbioe.2022.848699
Int J Mol Sci. 2022 Feb 25. pii: 2549. [Epub ahead of print]23(5):

Alternative Brain Slice-on-a-Chip for Organotypic Culture and Effective Fluorescence Injection Testing.

Pedro Herreros, Silvia Tapia-González, Laura Sánchez-Olivares, María Fe Laguna Heras, Miguel Holgado.

  Mouse brain slices are one of the most common models to study brain development and functioning, increasing the number of study models that integrate microfluidic systems for hippocampal slice cultures. This report presents an alternative brain slice-on-a-chip, integrating an injection system inside the chip to dispense a fluorescent dye for long-term monitoring. Hippocampal slices have been cultured inside these chips, observing fluorescence signals from living cells, maintaining the cytoarchitecture of the slices. Having fluorescence images of biological samples inside the chip demonstrates the effectiveness of the staining process using the injection method avoiding leaks or biological contamination. The technology developed in this study presents a significant improvement in the local administration of reagents within a brain slice-on-a-chip system, which could be a suitable option for organotypic cultures in a microfluidic chip acting as a highly effective bioreactor.

Keywords:  brain slice; cell labelling; fluorescence imaging; microfluidics; organ-on-a-chip

DOI:  https://doi.org/10.3390/ijms23052549
Cell Regen. 2022 Mar 07. 11(1): 7

Brain organoid-on-chip system to study the effects of breast cancer derived exosomes on the neurodevelopment of brain.

Kangli Cui, Wenwen Chen, Rongkai Cao, Yingying Xie, Peng Wang, Yunsong Wu, Yaqing Wang, Jianhua Qin.

  Early human brain development can be affected by multiple prenatal factors that involve chemical exposures in utero, maternal health characteristics such as psychiatric disorders, and cancer. Breast cancer is one of the most common cancers worldwide arising pregnancy. However, it is not clear whether the breast cancer might influence the brain development of fetus. Exosomes secreted by breast cancer cells play a critical role in mediating intercellular communication and interplay between different organs. In this work, we engineered human induced pluripotent stem cells (hiPSCs)-derived brain organoids in an array of micropillar chip and probed the influences of breast cancer cell (MCF-7) derived-exosomes on the early neurodevelopment of brain. The formed brain organoids can recapitulate essential features of embryonic human brain at early stages, in terms of neurogenesis, forebrain regionalization, and cortical organization. Treatment with breast cancer cell derived-exosomes, brain organoids exhibited enhanced expression of stemness-related marker OCT4 and forebrain marker PAX6. RNA-seq analysis reflected several activated signaling pathways associated with breast cancer, medulloblastoma and neurogenesis in brain organoids induced by tumor-derived exosomes. These results suggested that breast cancer cell-derived exosomes might lead to the impaired neurodevelopment in the brain organoids and the carcinogenesis of brain organoids. It potentially implies the fetus of pregnant women with breast cancer has the risk of impaired neurodevelopmental disorder after birth.

Keywords:  Brain organoid; Breast cancer; Exosomes; Human induced pluripotent stem cell

DOI:  https://doi.org/10.1186/s13619-021-00102-7
Sci Rep. 2022 Mar 07. 12(1): 3622

Bordetella pertussis-infected innate immune cells drive the anti-pertussis response of human airway epithelium.

M M Kroes, A Miranda-Bedate, R H J Jacobi, E van Woudenbergh, G den Hartog, J P M van Putten, J de Wit, E Pinelli.

Pertussis is a severe respiratory tract infection caused by Bordetella pertussis. This bacterium infects the ciliated epithelium of the human airways. We investigated the epithelial cell response to B. pertussis infection in primary human airway epithelium (HAE) differentiated at air-liquid interface. Infection of the HAE cells mimicked several hallmarks of B. pertussis infection such as reduced epithelial barrier integrity and abrogation of mucociliary transport. Our data suggests mild immunological activation of HAE by B. pertussis indicated by secretion of IL-6 and CXCL8 and the enrichment of genes involved in bacterial recognition and innate immune processes. We identified IL-1β and IFNγ, present in conditioned media derived from B. pertussis-infected macrophage and NK cells, as essential immunological factors for inducing robust chemokine secretion by HAE in response to B. pertussis. In transwell migration assays, the chemokine-containing supernatants derived from this HAE induced monocyte migration. Our data suggests that the airway epithelium on its own has a limited immunological response to B. pertussis and that for a broad immune response communication with local innate immune cells is necessary. This highlights the importance of intercellular communication in the defense against B. pertussis infection and may assist in the rational design of improved pertussis vaccines.

DOI: https://doi.org/10.1038/s41598-022-07603-8
Tissue Eng Regen Med. 2022 Mar 08.

Cyclic Stretching Induces Maturation of Human-Induced Pluripotent Stem Cell-Derived Cardiomyocytes through Nuclear-Mechanotransduction.

Myeongjin Song, Yongjun Jang, Seung-Jong Kim, Yongdoo Park.

  BACKGROUND: During cardiogenesis, cardiac cells receive various stimuli, such as biomechanical and chemical cues, from the surrounding microenvironment, and these signals induce the maturation of heart cells. Mechanical force, especially tensile force in the heart, is one of the key stimuli that induce cardiomyocyte (CM) maturation through mechanotransduction, a process through which physical cues are transformed into biological responses. However, the effects and mechanisms of tensile force on cell maturation are poorly studied.METHODS: In this study, we developed a cyclic stretch system that mimics the mechanical environment of the heart by loading tensile force to human-induced pluripotent stem cell (hiPSC)-derived CMs. hiPSC-CMs cultured with the cyclic stretch system analyzed morphological change, immunofluorescent staining, expression of maturation markers in mRNA, and beating properties compared to static cultures.
RESULTS: hiPSC-CMs cultured with the cyclic stretch system showed increased cell alignment, sarcomere length and expression of maturation markers in mRNA, such as TNNI3, MYL2 and TTN, compared to static cultures. Especially, the expression of genes related to nuclear mechanotransduction, such as Yap1, Lamin A/C, plectin, and desmin, was increased in the cyclically stretched hiPSC-CMs. Furthermore, the volume of the nucleus was increased by as much as 120% in the cyclic stretch group.
CONCLUSION: These results revealed that nuclear mechanotransduction induced by tensile force is involved in CM maturation. Together, these findings provide novel evidence suggesting that nuclear mechanotransduction induced by tensile force is involved in the regulation of cardiac maturation.

Keywords:  Cardiomyocyte; Cyclic stretch; Maturation; Nuclear-mechanotransduction; Tensile force

DOI:  https://doi.org/10.1007/s13770-021-00427-z
Nat Chem Biol. 2022 Mar 10.

Organoid screening reveals epigenetic vulnerabilities in human colorectal cancer.

Kohta Toshimitsu, Ai Takano, Masayuki Fujii, Kazuhiro Togasaki, Mami Matano, Sirirat Takahashi, Takanori Kanai, Toshiro Sato.

Precision oncology presumes an accurate prediction of drug response on the basis of the molecular profile of tumors. However, the extent to which patient-derived tumor organoids recapitulate the response of in vivo tumors to a given drug remains obscure. To gain insights into the pharmacobiology of human colorectal cancer (CRC), we here created a robust drug screening platform for patient-derived colorectal organoids. Application of suspension culture increased organoid scalability, and a refinement of the culture condition enabled incorporation of normal and precursor organoids to high-throughput drug screening. Drug screening identified bromodomain and extra-terminal (BET) bromodomain protein inhibitor as a cancer-selective growth suppressor that targets genes aberrantly activated in CRC. A multi-omics analysis identified an association between checkpoint with forkhead and ring finger domaines (CHFR) silencing and paclitaxel sensitivity, which was further validated by gene engineering of organoids and in xenografts. Our findings highlight the utility of multiparametric validation in enhancing the biological and clinical fidelity of a drug screening system.

DOI: https://doi.org/10.1038/s41589-022-00984-x