Isolation, Purification, and Cultivation of Primary Retinal Microvascular Pericytes: A Novel Model Using Rats

Liu, Guanghui; Meng, Chun; Pan, Mingdong; Chen, Meng; Deng, Ran; Lin, Ling; Zhao, Li; Liu, Xiaoling

doi:10.1111/micc.12121

Cited by 15 publications

(15 citation statements)

References 71 publications

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“…In present study, pericytes uniformly expressed the cellular markers PDGFR- β , NG2 and desmin. Our results are consistent with Liu's study, who also proved these markers expressed in pericytes isolated from rats by mechanical morcellation and collagenase digestion [ 29 ].…”

Section: Discussionsupporting

confidence: 93%

“…All experiments were performed in accordance with the Research Ethics Committees of the People's Hospital, Peking University, China. We isolated primary rat retinal pericyte cells from the retinal microvessel of Sprague-Dawley (SD) rats, using a modified method of previously published articles [ 27 – 29 ]. Briefly, eyes from SD rats (4–6 weeks, 150–200 g) were incubated with cold Dulbecco's phosphate-buffered saline (PBS) containing penicillin-streptomycin antibiotic (500 U/mL) for 10 min.…”

Section: Methodsmentioning

confidence: 99%

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Apelin Protects Primary Rat Retinal Pericytes from Chemical Hypoxia-Induced Apoptosis

Chen

Tao

Feng

et al. 2015

Journal of Ophthalmology

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Pericytes are a population of cells that participate in normal vessel architecture and regulate permeability. Apelin, as the endogenous ligand of G protein-coupled receptor APJ, participates in a number of physiological and pathological processes. To date, the effect of apelin on pericyte is not clear. Our study aimed to investigate the potential protection mechanisms of apelin, with regard to primary rat retinal pericytes under hypoxia. Immunofluorescence staining revealed that pericytes colocalized with APJ in the fibrovascular membranes dissected from proliferative diabetic retinopathy patients. In the in vitro studies, we first demonstrated that the expression of apelin/APJ was upregulated in pericytes under hypoxia, and apelin increased pericytes proliferation and migration. Moreover, knockdown of apelin in pericyte was achieved via lentivirus-mediated RNA interference. After the inhibition of apelin, pericytes proliferation was inhibited significantly in hypoxia culture condition. Furthermore, exogenous recombinant apelin effectively prevented hypoxia-induced apoptosis through downregulating active-caspase 3 expression and increasing the ratio of B cell lymphoma-2 (Bcl-2)/Bcl-2 associated X protein (Bax) in pericytes. These results suggest that apelin suppressed hypoxia-induced pericytes injury, which indicated that apelin could be a potential therapeutic target for retinal angiogenic diseases.

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Section: Discussionsupporting

confidence: 93%

Section: Methodsmentioning

confidence: 99%

Apelin Protects Primary Rat Retinal Pericytes from Chemical Hypoxia-Induced Apoptosis

Chen

Tao

Feng

et al. 2015

Journal of Ophthalmology

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“…Microvessel pericytes were obtained from rat retina as previously described ( 13 ). Rat retinas were dissected as described above.…”

Section: Methodsmentioning

confidence: 99%

Blood-filled cerebrospinal fluid-enhanced pericyte microvasculature contraction in rat retina: A novel in vitro study of subarachnoid hemorrhage

Cui

et al. 2016

Experimental and Therapeutic Medicine

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Previously, it was widely accepted that the delayed ischemic injury and poor clinical outcome following subarachnoid hemorrhage (SAH) was caused by cerebral vasospasm. This classical theory was challenged by a clazosentan clinical trial, which failed to improve patient outcome, despite reversing angiographic vasospasm. One possible explanation for the results of this trial is the changes in microcirculation following SAH, particularly in pericytes, which are the primary cell type controlling microcirculation in the brain parenchyma. However, as a result of technical limitations and the lack of suitable models, there was no direct evidence of microvessel dysfunction following SAH. In the present study, whole-mount retinal microvasculature has been introduced to study microcirculation in the brain following experimental SAH in vitro. Artificial blood-filled cerebrospinal fluid (BSCF) was applied to the retinal microvasculature to test the hypothesis that the presence of subarachnoid blood affects the contractile properties of the pericytes containing cerebral microcirculation during the early phase of SAH. It was observed that BCSF induced retina microvessel contraction and that this contraction could be resolved by BCSF wash-out. Furthermore, BCSF application accelerated pericyte-populated collagen gel contraction and increased the expression of α-smooth muscle actin. In addition, BCSF induced an influx of calcium in cultured retinal pericytes. In conclusion, the present study demonstrates increased contractility of retinal microvessels and pericytes in the presence of BCSF in vitro. These findings suggest that pericyte contraction and microvascular dysfunction is induced following SAH, which could lead to greater susceptibility to SAH-induced ischemia.

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“…Desmin , a type III intermediate filament required for mechanical elasticity of contractile cells (38) expressed in pericytes (33), is also present in skeletal and smooth muscle cells (39). Chondroitin sulfate proteoglycan (nerve/glia antigen-2/ NG2) also called chondroitin sulfate proteoglycan 4, melanoma chondroitin sulfate proteoglycan and melanoma-associated chondroitin sulfate proteoglycan has multiple signaling and regulatory functions (40) including recruitment of small GTPases (41).…”

Section: Pericyte Localization Structure and Markersmentioning

confidence: 99%

“…Some studies suggest that pericytes are noncontractile, however through their transformation into smooth muscle cells (34, 81, 82) they progressively, through the intermediate phenotypes, acquire an ultimate contractile phenotype. However, many studies showed that in addition to noncontractile proteins the pericytes also contain several contractile proteins such as smooth muscle- and nonmuscle-specific isoforms of actin, myosin, tropomyosin (31,33, 83–85) and a cyclic guanosine monophosphate (cGMP)-dependent protein kinase that regulates muscle contraction (86). Recent real-time studies of pericytes and dynamics of capillary diameter in mice cortex using intra vital two-photon laser scanning microscopy showed unequivocally that pericytes are contractile and are able to modulate capillary blood flow in the brain (87).…”

Section: Pericyte Contractility and Blood Flow Facilitationmentioning

confidence: 99%

Pericytes, Microvasular Dysfunction, and Chronic Rejection

Kloc¹,

Kubiak²,

Li³

et al. 2015

Transplantation

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Chronic rejection of transplanted organs remains the main obstacle in the long-term success of organ transplantation. Thus, there is a persistent quest for development of anti-chronic rejection therapies and identification of novel molecular and cellular targets. One of the potential targets is the pericytes, the mural cells of microvessels, which regulate microvascular permeability, development and maturation by controlling endothelial cell functions and regulating tissue fibrosis and inflammatory response. In this review we discuss the potential of targeting pericytes in development of microvasular dysfunction and the molecular pathways involved in regulation of pericyte activities for anti-chronic rejection intervention.

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Isolation, Purification, and Cultivation of Primary Retinal Microvascular Pericytes: A Novel Model Using Rats

Abstract: Retinal microvascular pericytes can be readily obtained by our method. We report the first cultivation of primary RMPs from rats and establish a simple method for their isolation and purification.

Cited by 15 publications

References 71 publications

Apelin Protects Primary Rat Retinal Pericytes from Chemical Hypoxia-Induced Apoptosis

Apelin Protects Primary Rat Retinal Pericytes from Chemical Hypoxia-Induced Apoptosis

Blood-filled cerebrospinal fluid-enhanced pericyte microvasculature contraction in rat retina: A novel in vitro study of subarachnoid hemorrhage

Pericytes, Microvasular Dysfunction, and Chronic Rejection

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