High throughput interrogation of human liver stellate cells reveals microenvironmental regulation of phenotype

Brougham-Cook, Aidan; Jain, Ishita; Kukla, David A.; Masood, Faisal; Kimmel, Hannah R. C.; Ryoo, Hyeon; Khetani, Salman R.; Underhill, Gregory H.

doi:10.1016/j.actbio.2021.11.015

Cited by 16 publications

(17 citation statements)

References 55 publications

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“… 66–69 Additionally, it has been shown that endothelial cells exhibit higher levels of traction on the periphery of ECM islands compared to the center, as measured by traction force microscopy (TFM), while others have demonstrated that altering ECM composition and stiffness can impact traction force magnitudes. 35,43,70 Additionally, it has been reported that endothelial cell permeability is modulated by substrate stiffness through changes in cell-ECM traction stresses and cell–cell junctional tensions, with cells on soft substrates (1 kPa), promoting higher cell-cell junction integrity compared to stiffer substrates (11 kPa). 51 Our data support these findings and indicate that LSEC contractility can be modulated by microenvironmental context, and that relative LSEC contractility regulates LSEC phenotype.…”

Section: Discussionmentioning

confidence: 99%

Engineered matrix microenvironments reveal the heterogeneity of liver sinusoidal endothelial cell phenotypic responses

et al. 2022

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Fibrosis is one of the hallmarks of chronic liver disease and is associated with aberrant wound healing. Changes in the composition of the liver microenvironment during fibrosis result in a complex crosstalk of extracellular cues that promote altered behaviors in the cell types that comprise the liver sinusoid, particularly liver sinusoidal endothelial cells (LSECs). Recently, it has been observed that LSECs may sustain injury before other fibrogenesis-associated cells of the sinusoid, implicating LSECs as key actors in the fibrotic cascade. A high-throughput cellular microarray platform was used to deconstruct the collective influences of defined combinations of extracellular matrix (ECM) proteins, substrate stiffness, and soluble factors on primary human LSEC phenotype in vitro. We observed remarkable heterogeneity in LSEC phenotype as a function of stiffness, ECM, and soluble factor context. LYVE-1 and CD-31 expressions were highest on 1 kPa substrates, and the VE-cadherin junction localization was highest on 25 kPa substrates. Also, LSECs formed distinct spatial patterns of LYVE-1 expression, with LYVE-1+ cells observed in the center of multicellular domains, and pattern size regulated by microenvironmental context. ECM composition also influenced a substantial dynamic range of expression levels for all markers, and the collagen type IV was observed to promote elevated expressions of LYVE-1, VE-cadherin, and CD-31. These studies highlight key microenvironmental regulators of LSEC phenotype and reveal unique spatial patterning of the sinusoidal marker LYVE-1. Furthermore, these data provide insight into understanding more precisely how LSECs respond to fibrotic microenvironments, which will aid drug development and identification of targets to treat liver fibrosis.

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Section: Discussionmentioning

confidence: 99%

Engineered matrix microenvironments reveal the heterogeneity of liver sinusoidal endothelial cell phenotypic responses

et al. 2022

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“…Microarray Fabrication: Polyacrylamide (PA) hydrogels were prepared following previous protocols and detailed procedures can be found in the cited literature. [16,17,45] Briefly, 12 mm glass coverslips or standard glass microscopy slides were etched (0.2 N NaOH) and rinsed with deionized H 2 O. Subsequently, coverslips and slides were dried, silanized (2% v/v 3-(trimethoxysilyl)propyl methacrylate), washed with ethanol, and dried once more.…”

Section: Methodsmentioning

confidence: 99%

“…We first examined attachment patterns of human endometrial microvascular endothelial cells (HEMECs) on single and pairwise combinations of ten endometrial-inspired ECM biomolecules (Figure 1a; Collagens I, III, IV, and V; Fibronectin, Laminin, Lumican, Decorin, Hyaluronic Acid, and Decorin) on polyacrylamide gels within the same stiffness regime (≈6 kPa) of the endometrium (≈1-2 kPa). [15][16][17] The dehydrated polyacrylamide hydrogel substrate acted to entrap the ECM proteins without chemical modification; [18,19] thus, enabling independent tuning of the ECM composition and the mechanical properties defined by the underlying polyacrylamide substrate. A representative image of the whole microarrays with seeded HEMECs (DAPI stained nuclei) is shown in Figure 1b.…”

Section: Human Endometrial Microvascular Endothelial Cells Display Di...mentioning

confidence: 99%

Cell Chirality of Micropatterned Endometrial Microvascular Endothelial Cells

Zambuto,

Jain,

Theriault

et al. 2024

Adv Healthcare Materials

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Chirality is an intrinsic cellular property that describes cell polarization biases along the left‐right axis, apicobasal axis, or front‐rear axes. Cell chirality plays a significant role in the arrangement of organs in the body as well as the orientation of organelles, cytoskeletons, and cells. Vascular networks within the endometrium, the mucosal inner lining of the uterus, commonly display spiral architectures that rapidly form across the menstrual cycle. Herein, we systematically examine the role of endometrial‐relevant extracellular matrix stiffness, composition, and soluble signals on endometrial endothelial cell chirality using a high‐throughput microarray. Endometrial endothelial cells display marked patterns of chirality as individual cells and as cohorts in response to substrate stiffness and environmental cues. Vascular networks formed from endometrial endothelial cells also display shifts in chirality as a function of exogenous hormones. Changes in cellular‐scale chirality correlate with changes in vascular network parameters, suggesting a critical role for cellular chirality in directing endometrial vessel network organization.This article is protected by copyright. All rights reserved

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“…Polyacrylamide (PA) hydrogels were prepared following previous protocols 16, 17, 39 . Briefly, 12 mm glass coverslips and standard microscopy glass slides were etched by immersing them 0.2 N NaOH (Sigma-Aldrich 415413-1L) for 1 hour on an orbital shaker and then rinsing with dH2O.…”

Section: Methodsmentioning

confidence: 99%

Cell Chirality of Micropatterned Endometrial Microvascular Endothelial Cells

Zambuto,

Jain,

Theriault

et al. 2023

Preprint

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Chirality is an intrinsic cellular property that describes cell polarization biases along the left-right axis, apicobasal axis, or front-rear axes. Cell chirality plays a significant role in the arrangement of organs in the body as well as the orientation of organelles, cytoskeletons, and cells. Vascular networks within the endometrium, the mucosal inner lining of the uterus, commonly display spiral architectures that rapidly form across the menstrual cycle. Herein, we systematically examine the role of endometrial-relevant extracellular matrix stiffness, composition, and soluble signals on endometrial endothelial cell chirality using a high-throughput microarray. Endometrial endothelial cells display marked patterns of chirality as individual cells and as cohorts in response to substrate stiffness and environmental cues. Vascular networks formed from endometrial endothelial cells also display shifts in chirality as a function of exogenous hormones. Changes in cellular-scale chirality correlate with changes in vascular network parameters, suggesting a critical role for cellular chirality in directing endometrial vessel network organization.

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High throughput interrogation of human liver stellate cells reveals microenvironmental regulation of phenotype

Cited by 16 publications

References 55 publications

Engineered matrix microenvironments reveal the heterogeneity of liver sinusoidal endothelial cell phenotypic responses

Engineered matrix microenvironments reveal the heterogeneity of liver sinusoidal endothelial cell phenotypic responses

Cell Chirality of Micropatterned Endometrial Microvascular Endothelial Cells

Cell Chirality of Micropatterned Endometrial Microvascular Endothelial Cells

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