Impact of serum concentration in cell culture media on tight junction proteins within a multiorgan microphysiological system

Salih, Abdul Rahim Chethikkattuveli; Farooqi, Hafiz Muhammad Umer; Kim, Young Su; Lee, Sang Ho; Choi, Kyung Hyun

doi:10.1016/j.mee.2020.111405

Cited by 23 publications

(25 citation statements)

References 47 publications

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“…It was due to the ample supply of FBS within the microphysiological system by cell culture media. 10% FBS in cell culture media results in continuously increasing TEER values and tighter cell–cell tight junction formation compared to serum-free or serum reduced cell culture media [ 38 ]. TGF-β1 in the concentration of 5 ng/mL was used as a stimulant to induce fibrosis in the liver fibrosis-on-chip model [ 39 ].…”

Section: Resultsmentioning

confidence: 99%

Real-time monitoring of liver fibrosis through embedded sensors in a microphysiological system

et al. 2021

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Hepatic fibrosis is a foreshadowing of future adverse events like liver cirrhosis, liver failure, and cancer. Hepatic stellate cell activation is the main event of liver fibrosis, which results in excessive extracellular matrix deposition and hepatic parenchyma's disintegration. Several biochemical and molecular assays have been introduced for in vitro study of the hepatic fibrosis progression. However, they do not forecast real-time events happening to the in vitro models. Trans-epithelial electrical resistance (TEER) is used in cell culture science to measure cell monolayer barrier integrity. Herein, we explored TEER measurement's utility for monitoring fibrosis development in a dynamic cell culture microphysiological system. Immortal HepG2 cells and fibroblasts were co-cultured, and transforming growth factor β1 (TGF-β1) was used as a fibrosis stimulus to create a liver fibrosis-on-chip model. A glass chip-based embedded TEER and reactive oxygen species (ROS) sensors were employed to gauge the effect of TGF-β1 within the microphysiological system, which promotes a positive feedback response in fibrosis development. Furthermore, albumin, Urea, CYP450 measurements, and immunofluorescent microscopy were performed to correlate the following data with embedded sensors responses. We found that chip embedded electrochemical sensors could be used as a potential substitute for conventional end-point assays for studying fibrosis in microphysiological systems.

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Section: Resultsmentioning

confidence: 99%

Real-time monitoring of liver fibrosis through embedded sensors in a microphysiological system

et al. 2021

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“…The level of relevance of a microphysiological system (MPS) with human physiology makes them ideal for identifying the novel therapeutics. The organ-on-a-chip and spheroid culture systems are evolving to better mimic human pathophysiology, which is being enhanced by the integration of sensors and robotics [1][2][3][4][5][6][7][8]. Among MPS, spheroids are emerging with the potential to mimic human tissues in a 3D shape and function [9][10][11].…”

Section: Introductionmentioning

confidence: 99%

Gravity-Based Flow Efficient Perfusion Culture System for Spheroids Mimicking Liver Inflammation

Kim,

Asif,

Chethikkattuveli Salih

et al. 2021

Biomedicines

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The spheroid culture system provides an efficient method to emulate organ-specific pathophysiology, overcoming the traditional two-dimensional (2D) cell culture limitations. The intervention of microfluidics in the spheroid culture platform has the potential to enhance the capacity of in vitro microphysiological tissues for disease modeling. Conventionally, spheroid culture is carried out in static conditions, making the media nutrient-deficient around the spheroid periphery. The current approach tries to enhance the capacity of the spheroid culture platform by integrating the perfusion channel for dynamic culture conditions. A pro-inflammatory hepatic model was emulated using a coculture of HepG2 cell line, fibroblasts, and endothelial cells for validating the spheroid culture plate with a perfusable channel across the spheroid well. Enhanced proliferation and metabolic capacity of the microphysiological model were observed and further validated by metabolic assays. A comparative analysis of static and dynamic conditions validated the advantage of spheroid culture with dynamic media flow. Hepatic spheroids were found to have improved proliferation in dynamic flow conditions as compared to the static culture platform. The perfusable culture system for spheroids is more physiologically relevant as compared to the static spheroid culture system for disease and drug analysis.

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“…A previously validated TEER sensor was used as an additional parameter for assessing cell layer confluency in a chip platform [ 8 , 10 , 25 , 26 ]. The TEER sensor was used to assess the real-time effect of different ECMs on the barrier integrity of HepG2 cell monolayer formed in the MPS based on impedance monitoring for 144 h ( Figure 4 and Figure S9 ).…”

Section: Resultsmentioning

confidence: 99%

“…It was observed that ECM has a significant impact on the formation of tight junctions. TEER values among different ECMs have a substantial effect on the MPS-based real-time biological assays, as several researchers have employed TEER for estimating cell viability, fibrosis development, and FBS standardization [ 8 , 27 , 28 , 29 ]. It can be concluded that the choice of ECM is vital for developing the most physiologically relevant MPSs.…”

Section: Resultsmentioning

confidence: 99%

“…Issues concerning cell culture in MPSs, such as cell number, cell type, tissue-specific ECM, and standard biomarker testing methods, need to be standardized for emulating human physiology [ 7 ]. Salih et al studied the effect of serum concentration on tight junction protein within MPSs by using a TEER sensor, which highlighted the direct influence on tight junction proteins (TJPs) required for attachment and biomarker production [ 8 ]. Accumulating evidence has indicated the positive impact of MPS surfacing modification by ECM relevant for a specific tissue type [ 9 , 10 , 11 ].…”

Section: Introductionmentioning

confidence: 99%

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Extracellular Matrix Optimization for Enhanced Physiological Relevance in Hepatic Tissue-Chips

et al. 2021

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The cellular microenvironment is influenced explicitly by the extracellular matrix (ECM), the main tissue support biomaterial, as a decisive factor for tissue growth patterns. The recent emergence of hepatic microphysiological systems (MPS) provide the basic physiological emulation of the human liver for drug screening. However, engineering microfluidic devices with standardized surface coatings of ECM may improve MPS-based organ-specific emulation for improved drug screening. The influence of surface coatings of different ECM types on tissue development needs to be optimized. Additionally, an intensity-based image processing tool and transepithelial electrical resistance (TEER) sensor may assist in the analysis of tissue formation capacity under the influence of different ECM types. The current study highlights the role of ECM coatings for improved tissue formation, implying the additional role of image processing and TEER sensors. We studied hepatic tissue formation under the influence of multiple concentrations of Matrigel, collagen, fibronectin, and poly-L-lysine. Based on experimental data, a mathematical model was developed, and ECM concentrations were validated for better tissue development. TEER sensor and image processing data were used to evaluate the development of a hepatic MPS for human liver physiology modeling. Image analysis data for tissue formation was further strengthened by metabolic quantification of albumin, urea, and cytochrome P450. Standardized ECM type for MPS may improve clinical relevance for modeling hepatic tissue microenvironment, and image processing possibly enhance the tissue analysis of the MPS.

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Impact of serum concentration in cell culture media on tight junction proteins within a multiorgan microphysiological system

Cited by 23 publications

References 47 publications

Real-time monitoring of liver fibrosis through embedded sensors in a microphysiological system

Real-time monitoring of liver fibrosis through embedded sensors in a microphysiological system

Gravity-Based Flow Efficient Perfusion Culture System for Spheroids Mimicking Liver Inflammation

Extracellular Matrix Optimization for Enhanced Physiological Relevance in Hepatic Tissue-Chips

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