A Versatile Model of Microfluidic Perifusion System for the Evaluation of C-Peptide Secretion Profiles: Comparison Between Human Pancreatic Islets and HLSC-Derived Islet-Like Structures

Gomez, Yonathan; Navarro-Tableros, Víctor; Tetta, Ciro; Camussi, Giovanni; Brizzi, Maria Felice

doi:10.3390/biomedicines8020026

Cited by 7 publications

(3 citation statements)

References 24 publications

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“…The Puck microbioreactor was designed to achieve the same function as commercially available openable flow-through microfluidic devices. 18 , 19 Rather than purchasing clamps made from machined parts that use proprietary designs, we used commercially available lens tubes (SM1L05E; ThorLabs, Newton, NJ) and threaded retaining rings (SM1PRR; ThorLabs) as the basis of a cylindrical clamp. Multiple layers of microfluidic elements were laser-cut from preformed acrylic (8560K172; McMaster-Carr, Elmhurst, IL) and silicone (87315K71 and 87315K73; McMaster-Carr) sheets using an infrared laser cutter (Mini 30W; Epilog Laser, Golden, CO).…”

Section: Methodsmentioning

confidence: 99%

A Microfluidic Platform for the Time-Resolved Interrogation of Polarized Retinal Pigment Epithelial Cells

Spivey,

Yin,

Chaum

et al. 2023

Trans. Vis. Sci. Tech.

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Purpose Cells grown in milliliter volume devices have difficulty measuring low-abundance secreted factors due to low resulting concentrations. Using microfluidic devices increases concentration; however, the constrained geometry makes phenotypic characterization with transepithelial electrical resistance more difficult and less reliable. Our device resolves this problem. Methods We designed and built a novel microfluidic “Puck” assembly using laser-cut pieces from preformed sheets of silicone and commercial off-the-shelf parts. Transwell membranes containing polarized retinal pigment epithelial (RPE) cells were reversibly sealed within the Puck and used to study polarized protein secretion. Protein secretion from the apical and basal surfaces in response to hypoxic conditions was quantified using an immunoassay method. Computational fluid modeling was performed on the chamber design. Results Under hypoxic culture conditions (7% O 2 ), basal vascular endothelial growth factor (VEGF) secretion by polarized RPE cells increased significantly from 1.40 to 1.68 ng/mL over the first 2 hours ( P < 0.0013) and remained stably elevated through 4 hours. Conversely, VEGF secretion from the apical side remained constant under the same hypoxic conditions. Conclusions The Puck can be used to measure spatiotemporal protein secretion by polarized cells into apical and basal microniches in response to environmental conditions. Computational model results support the absence of biologically significant shear stress to the cells caused by the device. Translational Relevance The Puck can be used validate the mature phenotypic health of autologous induced pluripotent stem cells (iPSC)-derived RPE cells prior to transplantation.

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Section: Methodsmentioning

confidence: 99%

A Microfluidic Platform for the Time-Resolved Interrogation of Polarized Retinal Pigment Epithelial Cells

Spivey,

Yin,

Chaum

et al. 2023

Trans. Vis. Sci. Tech.

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“…Under basal conditions, HLSC-ILSs synthesized and secreted basal levels of insulin and human C-peptide exhibiting an immature phenotype ( Navarro-Tableros et al, 2019 ). However, in the presence of high glucose and potassium concentrations, HLSC-ILSs were able to increase the levels of secreted C-peptide ( Navarro-Tableros et al, 2019 ; Gomez et al, 2020 ). In an in vivo setting, when implanted under the kidney capsule of streptozotocin-diabetic immunodeficient mice, HLSC-ILSs significantly reduced hyperglycemia.…”

Section: Hlsc Differentiation Abilities In Vitromentioning

confidence: 99%

Human Liver Stem Cells: A Liver-Derived Mesenchymal Stromal Cell-Like Population With Pro-regenerative Properties

Bruno¹,

Sanchez²,

Chiabotto³

et al. 2021

Front. Cell Dev. Biol.

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Human liver stem cells (HLSCs) were described for the first time in 2006 as a new stem cell population derived from healthy human livers. Like mesenchymal stromal cells, HLSCs exhibit multipotent and immunomodulatory properties. HLSCs can differentiate into several lineages under defined in vitro conditions, such as mature hepatocytes, osteocytes, endothelial cells, and islet-like cell organoids. Over the years, HLSCs have been shown to contribute to tissue repair and regeneration in different in vivo models, leading to more than five granted patents and over 15 peer reviewed scientific articles elucidating their potential therapeutic role in various experimental pathologies. In addition, HLSCs have recently completed a Phase 1 study evaluating their safety post intrahepatic injection in infants with inherited neonatal onset hyperammonemia. Even though a lot of progress has been made in understanding HLSCs over the past years, some important questions regarding the mechanisms of action remain to be elucidated. Among the mechanisms of interaction of HLSCs with their environment, a paracrine interface has emerged involving extracellular vesicles (EVs) as vehicles for transferring active biological materials. In our group, the EVs derived from HLSCs have been studied in vitro as well as in vivo. Our attention has mainly been focused on understanding the in vivo ability of HLSC–derived EVs as modulators of tissue regeneration, inflammation, fibrosis, and tumor growth. This review article aims to discuss in detail the role of HLSCs and HLSC-EVs in these processes and their possible future therapeutic applications.

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“…Insulin secretion is by far the most popular islet output and can be measured in real time or in downstream off‐chip analysis. Insulin secretion itself can be indicative of islet health if isolated from patients with diabetes 75 . Continuous real‐time on‐chip measurements of insulin output can be taken using fluorescence anisotropy from islet arrays 62 or single islets 76 .…”

Section: Engineered Fluidic Platforms For Studying Islets and Insulinmentioning

confidence: 99%

Engineering-inspired approaches to study β-cell function and diabetes

Lewis

Wells

2021

Stem Cells

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Strategies to mitigate the pathologies from diabetes range from simply administering insulin to prescribing complex drug/biologic regimens combined with lifestyle changes. There is a substantial effort to better understand β-cell physiology during diabetes pathogenesis as a means to develop improved therapies. The convergence of multiple fields ranging from developmental biology to microfluidic engineering has led to the development of new experimental systems to better study complex aspects of diabetes and β-cell biology. Here we discuss the available insulin-secreting cell types used in research, ranging from primary human β-cells, to cell lines, to pluripotent stem cell-derived β-like cells. Each of these sources possess inherent strengths and weaknesses pertinent to specific applications, especially in the context of engineered platforms. We then outline how insulin-expressing cells have been used in engineered platforms and how recent advances allow for better mimicry of in vivo conditions. Chief among these conditions are β-cell interactions with other endocrine organs. This facet is beginning to be thoroughly addressed by the organ-on-a-chip community, but holds enormous potential in the development of novel diabetes therapeutics. Furthermore, high throughput strategies focused on studying β-cell biology, improving β-cell differentiation, or proliferation have led to enormous contributions in the field and will no doubt be instrumental in bringing new diabetes therapeutics to the clinic.

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A Versatile Model of Microfluidic Perifusion System for the Evaluation of C-Peptide Secretion Profiles: Comparison Between Human Pancreatic Islets and HLSC-Derived Islet-Like Structures

Cited by 7 publications

References 24 publications

A Microfluidic Platform for the Time-Resolved Interrogation of Polarized Retinal Pigment Epithelial Cells

A Microfluidic Platform for the Time-Resolved Interrogation of Polarized Retinal Pigment Epithelial Cells

Human Liver Stem Cells: A Liver-Derived Mesenchymal Stromal Cell-Like Population With Pro-regenerative Properties

Engineering-inspired approaches to study β-cell function and diabetes

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