Isolation, cultivation, and characterization of primary bovine cochlear pericytes: A new in vitro model of stria vascularis

Abstract

The study of strial pericytes has gained great interest as they are pivotal for the physiology of stria vascularis. To provide an easily accessible in vitro model, here we described a growth medium‐based approach to obtain and cultivate primary bovine cochlear pericytes (BCP) from the stria vascularis of explanted bovine cochleae. We obtained high‐quality pericytes in 8–10 days with a > 90% purity after the second passage. Immunocytochemical analysis showed a homogeneous population of cells expressing typical pericyte markers, such as neural/glial antigen 2 (NG2), platelet‐derived growth factor receptorβ (PDGFRβ), α‐smooth muscle actin (α‐SMA), and negative for the endothelial marker von Willebrand factor. When challenged with tumor necrosis factor or lipopolysaccharide, BCP changed their shape, similarly to human retinal pericytes (HRPC). The sensitivity of BCP to ototoxic drugs was evaluated by challenging with cisplatin or gentamicin for 48 hr. Compared to human retinal endothelial cells and HRPC, cell viability of BCP was significantly lower ( p < 0.05) after the treatment with gentamicin or cisplatin. These data indicate that our protocol provides a simple and reliable method to obtain highly pure strial BCP. Furthermore, BCP are suitable to assess the safety profile of molecules which supposedly exert ototoxic activity, and may represent a valid alternative to in vivo tests.

Lipopolysaccharide decreases cochlear blood flow dose dependently in a guinea pig animal model via TNF signaling

“…TNF has been shown to cause cochlear pericytes to contract and thus reduce cochlear blood flow 34 . Additionally, LPS may directly affect cochlear pericytes, as has been observed in different tissues 35 as well as in cochlear pericytes after TNF exposure in vitro 36 …”

“…34 Additionally, LPS may directly affect cochlear pericytes, as has been observed in different tissues 35 as well as in cochlear pericytes after TNF exposure in vitro. 36 It has also been shown that in the central nervous system, pericytes are among the first cells to sustain damage in persistent hypoxia 37,38 and-if hypoxia persists long enough-may enter a rigor mortis like state, thus persistently impairing capillary blood flow. 39 Considering the similar physiological properties, a similar mechanism seems well probable.…”

Activation of the VEGF-A/ERK/PLA2 Axis Mediates Early Retinal Endothelial Cell Damage Induced by High Glucose: New Insight from an In Vitro Model of Diabetic Retinopathy

“…Primary human retinal endothelial cells (HRECs) were purchased from Innoprot (Elexalde Derio, Spain) and were fed with culture endothelial cell medium ECM, supplemented with 5% fetal bovine serum (FBS), 1% endothelial cell growth supplement (ECGS), 100 U/mL penicillin, and 100µg/mL streptomycin, provided by Innoprot. HRECs were characterized by their positive immunostaining for von Willebrand factor [ 63 ]. Cells were seeded in flasks or dishes pre-coated with a poly-L-Lysine solution (PLL), at a concentration of 1 mg/mL (Innoprot), for 1 h at 37 °C in a humidified atmosphere of 5% CO2.…”

Retinal ischemia induces α-SMA-mediated capillary pericyte contraction coincident with perivascular glycogen depletion

“…Pericytes with contractile capacity have been identified in several microvascular beds in addition to the retina and brain, where their role in maintaining blood retina/brain barrier and regulating neurovascular coupling at the microcirculatory level have been well-documented [12]. Similarly, cochlear pericytes also express α-SMA [13] and are thought to be involved in regulation of cochlear microcirculation [14, 15]. Renal pericytes as well control medullary blood flow by Ca 2+ -depended contractions [16].…”