Regulation of valve endothelial cell vasculogenic network architectures with ROCK and Rac inhibitors

Arevalos, Christopher; Walborn, Amanda; Rupert, Amanda A.; Berg, Jonathan M.; Godfrey, Elizabeth L.; Nguyen, Jacqueline; Grande-Allen, K. Jane

doi:10.1016/j.mvr.2015.01.008

Cited by 2 publications

(5 citation statements)

References 71 publications

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“…6 This effect of ROCK inhibition on network size of VIC-VEC co-cultures is consistent with previous reports that ROCK inhibition increases vasculogenic network size in VEC-only cultures. 1 Given that ROCK also regulates contraction of collagen gels by VICs, 10 this data suggests that early VIC-mediated VEC network organization is ROCK dependent. Furthermore, there is recent evidence that VICs can regulate the process of endothelial to mesenchyme transformation in VECs 14,28 ; a process that is also ROCK-dependent 27 .…”

Section: Discussionmentioning

confidence: 98%

“…To investigate the role of ROCK signaling on VEC/VIC network organization, once seeded the co-cultures were treated with the small molecule ROCK inhibitor Y-27632 (Calbiochem, San Diego, CA) 1 .…”

Section: Pkh Co-culture Tubule-like Structure Matrigel Assaymentioning

confidence: 99%

“…1 The total network length and number of tubule like structures were analyzed, and the resultant average tubule length was calculated using equation (1):…”

Section: Automated Network Analysismentioning

confidence: 99%

“…After 7 hours of co-culture, VECs cultured alone were organized ( Figure 1A) as previously reported. 1 PKH labeling of VECs (green) and VICs (red) in co-culture demonstrated that they organized together in well-defined networks with VICs scattered among the nodes and along the length of the tubule-like structures ( Figure 1B). This arrangement was similar to previous reports of PKH-tracked vascular ECs and pericytes (PCs).…”

Section: Vic-driven Collapse Of Vec Vasculogenic Network Is Rock-depmentioning

confidence: 99%

“…TX). VECs and VICs were harvested and cultured from the aortic valves as previously described 1,32 , and used between passages 2-4.…”

Section: Introductionmentioning

confidence: 99%

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Valve Interstitial Cells Act in a Pericyte Manner Promoting Angiogensis and Invasion by Valve Endothelial Cells

et al. 2016

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Neovascularization is an understudied aspect of calcific aortic valve disease (CAVD). Within diseased valves, cells along the neovessels' periphery stain for pericyte markers, but it is unclear whether valvular interstitial cells (VICs) can demonstrate a pericyte-like phenotype. This investigation examined the perivascular potential of VICs to regulate valve endothelial cell (VEC) organization and explored the role of Angiopoeitin1-Tie2 signaling in this process. Porcine VECs and VICs were fluorescently tracked and co-cultured in Matrigel over 7 days. VICs regulated early VEC network organization in a ROCK-dependent manner, then guided later VEC network contraction through chemoattraction. Unlike vascular control cells, the valve cell cultures ultimately formed invasive spheroids with 3D angiogenic-like sprouts. VECs co-cultured with VICs displayed significantly more invasion than VECs alone; with VICs generally leading and wrapping around VEC invasive sprouts. Lastly, Angiopoietin1-Tie2 signaling was found to regulate valve cell organization during VEC/VIC spheroid formation and invasion. VICs demonstrated pericyte-like behaviors toward VECs throughout sustained co-culture. The change from a vasculogenic network to an invasive sprouting spheroid suggests that both cell types undergo phenotypic changes during long-term culture in the model angiogenic environment. Valve cells organizing into spheroids and undergoing 3D invasion of Matrigel demonstrated several typical angiogenic-like phenotypes dependent on basal levels of Angiopoeitin1-Tie2 signaling and ROCK activation. These results suggest that the ectopic sustained angiogenic environment during the early stages of valve disease promotes organized activity by both VECs and VICs, contributing to neovessel formation and the progression of CAVD.

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Section: Discussionmentioning

confidence: 98%

Section: Pkh Co-culture Tubule-like Structure Matrigel Assaymentioning

confidence: 99%

“…1 The total network length and number of tubule like structures were analyzed, and the resultant average tubule length was calculated using equation (1):…”

Section: Automated Network Analysismentioning

confidence: 99%

Section: Vic-driven Collapse Of Vec Vasculogenic Network Is Rock-depmentioning

confidence: 99%

“…TX). VECs and VICs were harvested and cultured from the aortic valves as previously described 1,32 , and used between passages 2-4.…”

Section: Introductionmentioning

confidence: 99%

See 3 more Smart Citations