Transcriptional profiles of valvular interstitial cells cultured on tissue culture polystyrene, on 2D hydrogels, or within 3D hydrogels

Mabry, Kelly M.; Payne, Samuel Z.; Anseth, Kristi S.

doi:10.1016/j.dib.2015.11.017

Cited by 4 publications

(6 citation statements)

References 5 publications

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“…mRNA expression levels in VICs cultured on TCPS, 2D hydrogels, or within 3D hydrogels were compared to freshly isolated VICs using a porcine DNA microarray [23]. VICs cultured on TCPS had differential expression compared to freshly isolated cells for 3304 probesets (out of 25470), where a difference was defined as a fold change greater than 2 or less than −2 and a p-value less than 0.05 to achieve statistical and biological significance (Figure 3A).…”

Section: Resultsmentioning

confidence: 99%

Microarray analyses to quantify advantages of 2D and 3D hydrogel culture systems in maintaining the native valvular interstitial cell phenotype

2016

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Valvular interstitial cells (VICs) actively maintain and repair heart valve tissue; however, persistent activation of VICs to a myofibroblast phenotype can lead to aortic stenosis. To better understand and quantify how microenvironmental cues influence VIC phenotype and myofibroblast activation, we compared expression profiles of VICs cultured on poly(ethylene glycol) (PEG) gels to those cultured on tissue culture polystyrene (TCPS), as well as fresh isolates. In general, VICs cultured in hydrogel matrices had lower levels of activation (<10%), similar to levels seen in healthy valve tissue, while VICs cultured on TCPS were ~75% activated myofibroblasts. VICs cultured on TCPS also exhibited a higher magnitude of perturbations in gene expression than soft hydrogel cultures when compared to the native phenotype. Using peptide-modified PEG gels, VICs were seeded on (2D), as well as encapsulated in (3D), matrices of the same composition and modulus. Despite similar levels of activation, VICs cultured in 2D had distinct variations in transcriptional profiles compared to those in 3D hydrogels. Genes related to cell structure and motility were particularly affected by the dimensionality of the culture platform, with higher expression levels in 2D than in 3D. These results indicate that dimensionality may play a significant role in dictating cell phenotype (e.g., through differences in polarity, diffusion of soluble signals), and emphasize the importance of using multiple metrics when characterizing cell phenotype.

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Section: Resultsmentioning

confidence: 99%

Microarray analyses to quantify advantages of 2D and 3D hydrogel culture systems in maintaining the native valvular interstitial cell phenotype

2016

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Section: Introductionmentioning

confidence: 99%

“…By now, various in vitro CAVD models have been described in the literature [3][4][5][6][7]. Beside conventional 2D cell culture, 3D models have recently become increasingly popular [6][7][8][9][10]. These 3D models are more likely to rebuild the actual cell and ECM interactions of the native valve [11][12][13][14].…”

Section: Introductionmentioning

confidence: 99%

3D-bioprinting of aortic valve interstitial cells: impact of hydrogel and printing parameters on cell viability

et al. 2022

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Background: Calcific aortic valve disease (CAVD) is a frequent cardiac pathology in the aging society. Although valvular interstitial cells (VIC) seem to play a crucial role, mechanisms of CAVD are not fully understood. Development of tissue-engineered cellular models by 3D-bioprinting may help to further investigate underlying mechanisms of CAVD. Methods: VIC were isolated from ovine aortic valves and cultured in Dulbecco’s Modified Eagle’s Medium (DMEM). VIC of passages six to ten were dissolved in a hydrogel consisting of 2% alginate and 8% gelatin with a concentration of 2x106 VIC/ml. Cell-free and VIC-laden hydrogels were printed with an extrusion-based 3D-bioprinter (3D-Bioplotter® Developer Series, EnvisionTec, Gladbeck, Germany), cross-linked and incubated for up to 28 days. Accuracy and durability of scaffolds was examined by microscopy and cell viability was tested by cell counting kit-8 assay and live/dead staining. Results: 3D-bioprinting of scaffolds was most accurate with a printing pressure of P<400 hPa, nozzle speed of v<20 mm/s, hydrogel temperature of TH=37 °C and platform temperature of TP=5 °C in a 90° parallel line as well as in a honeycomb pattern. Dissolving the hydrogel components in DMEM increased VIC viability on day 21 by 2.5-fold compared to regular 0.5% saline-based hydrogels (p<0.01). Examination at day 7 revealed dividing and proliferating cells. After 21 days the entire printed scaffolds were filled with proliferating cells. Live/dead cell viability/cytotoxicity staining confirmed beneficial effects of DMEM-based cell-laden VIC hydrogel scaffolds even 28 days after printing. Conclusions: By using low pressure printing methods, we were able to successfully culture cell-laden 3D-bioprinted VIC scaffolds for up to 28 days. Using DMEM-based hydrogels can significantly improve the long-term cell viability and overcome printing-related cell damage. Therefore, future applications 3D-bioprinting of VIC might enable the development of novel tissue engineered cellular 3D-models to examine mechanisms involved in initiation and progression of CAVD.

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“…The phenotype of valvular interstitial cells (VICs), the most abundant and plastic cell population in the valve, can be modulated by the properties of cell culture substrate/matrix . It has been revealed that matrix stiffness affects the activation and osteogenic differentiation of VICs . Moreover, the morphology and architecture of valvular ECM have been shown to instruct the attachment and spreading of VICs .…”

Section: Introductionmentioning

confidence: 99%

“…6,7 It has been revealed that matrix stiffness affects the activation and osteogenic differentiation of VICs. [8][9][10] Moreover, the morphology and architecture of valvular ECM have been shown to instruct the attachment and spreading of VICs. 11,12 Several studies have performed uniaxial tensile testing on tissue-engineered scaffolds, 13,14 but uniaxial testing is insufficient to fully characterize the scaffold mechanical properties as it fails to capture the complex cross-coupling (stress along one axis is affected by the strain level along the perpendicular axis) experienced by the anisotropic tissues.…”

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confidence: 99%

Valve leaflet‐inspired elastomeric scaffolds with tunable and anisotropic mechanical properties

Xue

Ravishankar

Zeballos

et al. 2019

Polymers for Advanced Techs

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Fibrous scaffolds, which can mimic the elastic and anisotropic mechanical properties of native tissues, hold great promise in recapitulating the native tissue microenvironment. We previously fabricated electrospun fibrous scaffolds made of hybrid synthetic elastomers (poly(1,3‐diamino‐2‐hydroxypropane‐co‐glycerol sebacate)‐co‐poly (ethylene glycol) (APS‐co‐PEG) and polycaprolactone (PCL)) to obtain uniaxial mechanical properties similar to those of human aortic valve leaflets. However, conventional electrospinning process often yields scaffolds with random alignment, which fails to recreate the anisotropic nature of most of the soft tissues such as native heart valves. Inspired by the structure of native valve leaflet, we designed a novel valve leaflet‐inspired ring‐shaped collector to modulate the electrospun fiber alignment and studied the effect of polymer formulation (PEG amount [mole %] in APS‐co‐PEG; ratio between APS‐co‐PEG and PCL; and total polymer concentration) in tuning the biaxial mechanical properties of the fibrous scaffolds. The fibrous scaffolds collected on the ring‐shaped collector displayed anisotropic biaxial mechanical properties, suggesting that their biaxial mechanical properties are closely associated with the fiber alignment in the scaffold. Additionally, the scaffold stiffness was easily tuned by changing the composition and concentration of the polymer blend. Human valvular interstitial cells (hVICs) cultured on these anisotropic scaffolds displayed aligned morphology as instructed by the fiber alignment. Overall, we generated a library of biologically relevant fibrous scaffolds with tunable mechanical properties, which will guide the cellular alignment.

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Transcriptional profiles of valvular interstitial cells cultured on tissue culture polystyrene, on 2D hydrogels, or within 3D hydrogels

Cited by 4 publications

References 5 publications

Microarray analyses to quantify advantages of 2D and 3D hydrogel culture systems in maintaining the native valvular interstitial cell phenotype

Microarray analyses to quantify advantages of 2D and 3D hydrogel culture systems in maintaining the native valvular interstitial cell phenotype

3D-bioprinting of aortic valve interstitial cells: impact of hydrogel and printing parameters on cell viability

Valve leaflet‐inspired elastomeric scaffolds with tunable and anisotropic mechanical properties

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