Shear stress upregulates regeneration‐related immediate early genes in liver progenitors in 3D ECM‐like microenvironments

Nishii, Kenichiro; Brodin, Erik; Renshaw, Taylor; Weesner, Rachael; Moran, Emma; Söker, Shay; Sparks, Jessica L.

doi:10.1002/jcp.26246

Cited by 20 publications

(15 citation statements)

References 49 publications

(85 reference statements)

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“…While static/cyclic stretches or fluid stresses were previously shown to induce a transient increase in IEG transcript levels ( 17 , 18 , 40 ), we cannot completely rule out the possibility that trypsinization could represent a significant mechanical strain. However, it is worth noting that: (i) the same trypsinization procedure was applied to all samples in the time course experiment, (ii) only a small fraction of gonadotrope cells (<10%) exhibited IEG activation at the 0 and 0.5 nM GnRH concentrations, (iii) the vast majority of cells (>90%) were activated at the 20 nM concentration.…”

Section: Discussionmentioning

confidence: 77%

“…Dissecting immediate-early transcriptional responses to an external stimulus in individual cells requires elaborate study designs that include various treatments, time points, and replicates. IEGs, in particular, are dynamic and can respond to handling ( 17 , 18 ). The use of fresh cells for SC transcriptomics necessitates immediate sample processing, which is incompatible with complex time course studies and with a separation between sample harvest and SC processing steps.…”

Section: Introductionmentioning

confidence: 99%

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Single-cell stabilization method identifies gonadotrope transcriptional dynamics and pituitary cell type heterogeneity

Ruf-Zamojski

Nair

et al. 2018

Nucleic Acids Research

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Immediate-early response genes (IEGs) are rapidly and transiently induced following an extracellular signal. Elucidating the IEG response patterns in single cells (SCs) requires assaying large numbers of timed samples at high accuracy while minimizing handling effects. To achieve this, we developed and validated RNA stabilization Buffer for Examination of Single-cell Transcriptomes (RNA-Best), a versatile single-step cell and tissue preservation protocol that stabilizes RNA in intact SCs without perturbing transcription patterns. We characterize for the first time SC heterogeneity in IEG responses to pulsatile gonadotropin-releasing hormone (GnRH) stimuli in pituitary gonadotrope cells. Our study identifies a gene-specific hierarchical pattern of all-or-none transcript induction elicited by increasing concentrations of GnRH. This quantal pattern of gene activation raises the possibility that IEG activation, when accurately resolved at the SC level, may be mediated by gene bits that behave as pure binary switches.

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Section: Discussionmentioning

confidence: 77%

Section: Introductionmentioning

confidence: 99%

Single-cell stabilization method identifies gonadotrope transcriptional dynamics and pituitary cell type heterogeneity

Ruf-Zamojski

Nair

et al. 2018

Nucleic Acids Research

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“…Thus, fluid flow, shear stress in the microfluidic chips can affect the NSC biology. It is known that fluid mechanical stress resulted in differential gene expression compared to static control ( Nishii et al ., 2018 ) Thus, it is possible that NSCs in the microfluidic chamber may show different feature compare to the static culture.…”

Section: Discussionmentioning

confidence: 99%

Neural Stem Cell Differentiation Using Microfluidic Device-Generated Growth Factor Gradient

Kim

Sim

Kim

2018

Biomolecules & Therapeutics

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Neural stem cells (NSCs) have the ability to self-renew and differentiate into multiple nervous system cell types. During embryonic development, the concentrations of soluble biological molecules have a critical role in controlling cell proliferation, migration, differentiation and apoptosis. In an effort to find optimal culture conditions for the generation of desired cell types in vitro, we used a microfluidic chip-generated growth factor gradient system. In the current study, NSCs in the microfluidic device remained healthy during the entire period of cell culture, and proliferated and differentiated in response to the concentration gradient of growth factors (epithermal growth factor and basic fibroblast growth factor). We also showed that overexpression of ASCL1 in NSCs increased neuronal differentiation depending on the concentration gradient of growth factors generated in the microfluidic gradient chip. The microfluidic system allowed us to study concentration-dependent effects of growth factors within a single device, while a traditional system requires multiple independent cultures using fixed growth factor concentrations. Our study suggests that the microfluidic gradient-generating chip is a powerful tool for determining the optimal culture conditions.

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“…[ 366,368 ] Interestingly, recovery from acute liver injury or hepatectomy is also dependent on increases in shear stress and flow, which induces release of NO from LSECs and HSCs and sensitizes hepatocytes to hepatocyte growth factor (HGF) necessary for proliferation and regeneration. [ 368,370 ]…”

Section: Modeling Vascular Mechanopathology In Vascularized Microphysmentioning

confidence: 99%

Biofabrication Strategies and Engineered In Vitro Systems for Vascular Mechanobiology

Pradhan

Banda

Farino

et al. 2020

Adv Healthcare Materials

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The vascular system is integral for maintaining organ‐specific functions and homeostasis. Dysregulation in vascular architecture and function can lead to various chronic or acute disorders. Investigation of the role of the vascular system in health and disease has been accelerated through the development of tissue‐engineered constructs and microphysiological on‐chip platforms. These in vitro systems permit studies of biochemical regulation of vascular networks and parenchymal tissue and provide mechanistic insights into the biophysical and hemodynamic forces acting in organ‐specific niches. Detailed understanding of these forces and the mechanotransductory pathways involved is necessary to develop preventative and therapeutic strategies targeting the vascular system. This review describes vascular structure and function, the role of hemodynamic forces in maintaining vascular homeostasis, and measurement approaches for cell and tissue level mechanical properties influencing vascular phenomena. State‐of‐the‐art techniques for fabricating in vitro microvascular systems, with varying degrees of biological and engineering complexity, are summarized. Finally, the role of vascular mechanobiology in organ‐specific niches and pathophysiological states, and efforts to recapitulate these events using in vitro microphysiological systems, are explored. It is hoped that this review will help readers appreciate the important, but understudied, role of vascular‐parenchymal mechanotransduction in health and disease toward developing mechanotherapeutics for treatment strategies.

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Shear stress upregulates regeneration‐related immediate early genes in liver progenitors in 3D ECM‐like microenvironments

Cited by 20 publications

References 49 publications

Single-cell stabilization method identifies gonadotrope transcriptional dynamics and pituitary cell type heterogeneity

Single-cell stabilization method identifies gonadotrope transcriptional dynamics and pituitary cell type heterogeneity

Neural Stem Cell Differentiation Using Microfluidic Device-Generated Growth Factor Gradient

Biofabrication Strategies and Engineered In Vitro Systems for Vascular Mechanobiology

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