Concomitant Evaluation of a Panel of Exosome Proteins and MiRs for Qualification of Cultured Human Corneal Endothelial Cells

Ueno, Masaki; Asada, Kazuko; Toda, Munetoyo; Nagata, Kazue; Sotozono, Chie; Kosaka, Nobuyoshi; Ochiya, Takahiro; Kinoshita, Shigeru; Hamuro, Junji

doi:10.1167/iovs.16-19805

Cited by 22 publications

(19 citation statements)

References 34 publications

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“…It should be noted that CD44 might be repressed by wild-type p53 29 in consistent with our previous finding of upregulated p53 in CD44−/+ differentiated SPs. 30 It should be noted that mutant p53 promotes aerobic glycolysis. 29 Given the recent finding that PDGF-ββ stimulated Glc uptake, lactate production, and ATP production through lactate dehydrogenase A (LDHA) 31 and the segregated upregulation of LDHA in CD44++/+++ SPs (Fig.…”

Section: Discussionmentioning

confidence: 99%

“…Some of the observations were fully consistent with our previous observation describing the metabolomic profiling segregated differentiated from dedifferentiated-cHCECs, 9 the linkage of the dedifferentiation state with epithelial-mesenchymal transition and transformed fibroblastic cell morphology, 7,8 and the observation that cHCECs SPs with CST secreted higher amount of exosomes than the mature differentiated cHCECs SPs. 30…”

Section: Integral Proteome Analysis: Gene Ontology Analysismentioning

confidence: 99%

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Metabolites Interrogation in Cell Fate Decision of Cultured Human Corneal Endothelial Cells

Hamuro

Numa

Fujita

et al. 2020

Invest. Ophthalmol. Vis. Sci.

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Aiming to clarify the metabolic interrogation in cell fate decision of cultured human corneal endothelial cells (cHCECs). METHODS. To analyze the metabolites in the culture supernatants (CS), 34 metabolome measurements were carried out for mature differentiated and a variety of cHCECs with cell state transition through a facility service. Integrated proteomics research for cell lysates by liquid chromatography−tandem mass spectrometry (LC-MS/MS) was performed for 3 aliquots of each high-quality or low-quality cHCEC subpopulations (SP). The investigations for the focused genes involved in cHCEC metabolism were performed by using DAVID and its options "KEGG_PATHWAY." RESULTS. The clusters of metabolites coincided well with the distinct content of CD44−/+ SPs. Both secreted pyruvic acid and lactic acid in the CS were negatively correlated with the content of high-quality SPs. Lactic acid and pyruvic acid in the CS exhibited the positive correlation with that of Ile, Leu, and Ser, whereas the negative correlation was with glutamine. Platelet-derived growth factor-ββ in the CS negatively correlated with lactic acid in CS, indicating indirectly the positive correlation with the content of CD44−/+ SPs. Upregulated glycolytic enzymes and influx of glutamine to the tricarboxylic acid cycle may be linked with a metabolic rewiring converting oxidative metabolism in mature differentiated CD44−/+SPs into a glycolytic flux-dependent state in immature SPs with cell state transition. CONCLUSIONS. The findings suggest that the cell fate decision of cHCECs may be dictated at least partly through metabolic rewiring.

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Section: Discussionmentioning

confidence: 99%

Section: Integral Proteome Analysis: Gene Ontology Analysismentioning

confidence: 99%

Metabolites Interrogation in Cell Fate Decision of Cultured Human Corneal Endothelial Cells

Hamuro

Numa

Fujita

et al. 2020

Invest. Ophthalmol. Vis. Sci.

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“…For the purposes of this review, we will consider the blood–spinal cord barrier in the same context as the BBB, though subtle differences between the two have been noted [ 27 ]. The blood–cerebrospinal barrier (BCSFB) is comprised of epithelial cells of the choroid plexus (localized in the brain ventricles) that separate blood–borne elements from the cerebrospinal fluid (CSF) [ 28 ]. A third type CNS barrier, the blood–leptomeningeal barrier (BLMB), is formed by a monolayer of endothelial cells of the microvessels coursing through the pia mater and overlying subarachnoid space (SAS), and may be considered another type of BCSFB [ 29 ].…”

Section: Introductionmentioning

confidence: 99%

Extracellular vesicles: mediators and biomarkers of pathology along CNS barriers

Ramirez

Andrews

Paul

et al. 2018

Fluids Barriers CNS

119

108

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Extracellular vesicles (EVs) are heterogeneous, nano-sized vesicles that are shed into the blood and other body fluids, which disperse a variety of bioactive molecules (e.g., protein, mRNA, miRNA, DNA and lipids) to cellular targets over long and short distances. EVs are thought to be produced by nearly every cell type, however this review will focus specifically on EVs that originate from cells at the interface of CNS barriers. Highlighted topics include, EV biogenesis, the production of EVs in response to neuroinflammation, role in intercellular communication and their utility as a therapeutic platform. In this review, novel concepts regarding the use of EVs as biomarkers for BBB status and as facilitators for immune neuroinvasion are also discussed. Future directions and prospective are covered along with important unanswered questions in the field of CNS endothelial EV biology.

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“…SLC4A11 is a highly expressed transporter protein in the corneal endothelium that plays a significant role in CEnC function, and biallelic mutation of this protein leads to congenital hereditary endothelial dystrophy (Aldave et al, 2013; Jiao et al, 2007; Vithana et al, 2006). Conversely, CD44 is not expressed in the corneal endothelium, and its expression correlates with a senescent phenotype in vascular and corneal endothelial cells (Mun and Boo, 2010; Ueno et al, 2016).…”

Section: Discussionmentioning

confidence: 99%

Phenotypic and functional characterization of corneal endothelial cells during in vitro expansion

Frausto¹,

Swamy²,

Peh

et al. 2019

Preprint

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SUMMARYThe advent of cell culture-based methods for the establishment and expansion of human corneal endothelial cells (CEnC) has provided a source of transplantable corneal endothelium, with a significant potential to challenge the one donor-one recipient paradigm. However, concerns over cell identity remain, and a comprehensive characterization of the cultured CEnC across serial passages has not been performed. To this end, we compared two established CEnC culture methods by assessing the transcriptomic changes that occur during in vitro expansion. In confluent monolayers, low mitogenic culture conditions preserved corneal endothelial cell state identity better than culture in high mitogenic conditions. Expansion by continuous passaging induced replicative cell senescence. Transcriptomic analysis of the senescent phenotype identified a cell senescence signature distinct for CEnC. We identified activation of both classic and new cell signaling pathways that may be targeted to prevent senescence, a significant barrier to realizing the potential clinical utility of in vitro expansion.

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Concomitant Evaluation of a Panel of Exosome Proteins and MiRs for Qualification of Cultured Human Corneal Endothelial Cells

Cited by 22 publications

References 34 publications

Metabolites Interrogation in Cell Fate Decision of Cultured Human Corneal Endothelial Cells

Metabolites Interrogation in Cell Fate Decision of Cultured Human Corneal Endothelial Cells

Extracellular vesicles: mediators and biomarkers of pathology along CNS barriers

Phenotypic and functional characterization of corneal endothelial cells during in vitro expansion

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