Isolation of Endothelial Cells from Brain, Lung, and Kidney: Expression of the Multidrug Resistance P-Glycoprotein Isoforms

Demeule, Michel; Labelle, Myriam; Régina, Anthony; Béliveau, Richard

doi:10.1006/bbrc.2001.4312

Cited by 99 publications

(61 citation statements)

References 38 publications

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“…Lung and bone marrow SP cells also express the endothelial marker CD31. Several recent studies have suggested that endothelial cells and HSC are derived from a common origin (1,8,18,25). The coexpression of the hematopoietic marker CD45 and CD31 on lung SP cells further supports this link.…”

Section: Discussionmentioning

confidence: 60%

Side population cells and Bcrp1 expression in lung

Summer

Kotton

et al. 2003

American Journal of Physiology-Lung Cellular and Molecular Phys

242

196

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Side population (SP) cells are a rare subset of cells found in various tissues that are highly enriched for stem cell activity. SP cells can be isolated by dual-wavelength flow cytometry because of their capacity to efflux Hoechst dye, a process mediated by the ATP-binding cassette transporter breast cancer resistance protein (Bcrp) 1. By performing flow cytometry of enzymedigested mouse lung stained with Hoechst dye, we found that SP cells comprise 0.03–0.07% of total lung cells and are evenly distributed in proximal and distal lung regions. By RT-PCR, we found that lung SP cells express hepatocyte nuclear factor-3β, but not thyroid transcription factor-1. Surface marker analysis revealed lung SP cells to be stem cell antigen 1 positive, Bcrp1 positive, lineage marker negative, and heterogeneous at the CD45 locus. As expected, we did not detect lung SP cells in Bcrp1-deficient animals. We, therefore, employed nonisotopic in situ hybridization and immunostaining for Bcrp1 as a strategy to localize these cells in vivo. Expression was observed in distinct lung cell types: bronchial and vascular smooth muscle cells and round cells within the distal air space. We confirmed the expression of Bcrp1 in primary bronchial smooth muscle cell cultures (BSMC) and in lavaged distal airway cells, but neither possessed the capacity to efflux Hoechst dye. In BSMC, Bcrp1 was localized to an intracellular compartment, suggesting that the molecular site of Bcrp1 expression regulates SP phenotype.

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Section: Discussionmentioning

confidence: 60%

Side population cells and Bcrp1 expression in lung

Summer

Kotton

et al. 2003

American Journal of Physiology-Lung Cellular and Molecular Phys

242

196

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“…Glomerular endothelial cells were isolated with the method developed previously and were identified by positive immunofluorescence for the endothelial cell-specific marker CD31 (platelet/endothelial cell adhesion molecule-1). [22][23][24] Isolation and identification of rat primary tubular epithelial cells were performed according to previously published techniques, and cytokeratin 18 was used as a specific marker of tubular epithelial cells. [25][26][27] Interstitial fibroblasts were isolated and identified as reported.…”

Section: Isolation and Identification Of Primary Renal Residential Cellsmentioning

confidence: 99%

miR-335 and miR-34a Promote Renal Senescence by Suppressing Mitochondrial Antioxidative Enzymes

Bai

Ding

et al. 2011

Journal of the American Society of Nephrology

193

145

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The molecular basis for aging of the kidney is not well understood. MicroRNAs (miRNAs) contribute to processes such as development, differentiation, and apoptosis, but their contribution to the aging process is unknown. Here, we analyzed the miRNA expression profile of young (3-month) and old (24-month) rat kidneys and identified the biologic pathways and genes regulated by differentially expressed miRNAs. We observed upregulation of 18 miRNAs with aging, mainly regulating the genes associated with energy metabolism, cell proliferation, antioxidative defense, and extracellular matrix degradation; in contrast, we observed downregulation of 7 miRNAs with aging, principally targeting the genes associated with the immune inflammatory response and cell-cycle arrest. Bioinformatics analysis suggested that superoxide dismutase 2 (SOD2) and thioredoxin reductase 2 (Txnrd2), located in the mitochondria, are potential targets of miR-335 and miR-34a, respectively. Aging mesangial cells exhibited significant upregulation of miR-335 and miR-34a and marked downregulation of SOD2 and Txnrd2. miR-335 and miR-34a inhibited expression of SOD2 and Txnrd2 by binding to the 3Ј-untranslated regions of each gene, respectively. Overexpression of miR-335 and miR-34a induced premature senescence of young mesangial cells via suppression of SOD2 and Txnrd2 with a concomitant increase in reactive oxygen species (ROS). Conversely, antisense miR-335 and miR-34a inhibited senescence of old mesangial cells via upregulation of SOD2 and Txnrd2 with a concomitant decrease in ROS. In conclusion, these results suggest that miRNAs may contribute to renal aging by inhibiting intracellular pathways such as those involving the mitochondrial antioxidative enzymes SOD2 and Txnrd2. 22: 125222: -126122: , 201122: . doi: 10.1681 Kidney aging is an important clinical problem, not only because normal aging reduces renal function but also because of the high frequency of ESRD, renal cancer, and renal failure in elderly people. Renal aging is of interest as a general model for organ aging because renal function can be quantitatively assessed more readily than that of other organs in clinical practice. 1 At the present time, the molecular basis of renal aging is not clearly known. For example, nothing is known of the role of microRNAs (miRNAs) in the aging process of organs. J Am Soc NephrolmiRNAs are a novel class of small, regulatory, noncoding RNA molecules that inhibit the expression of multiple genes at the post-transcriptional level. miRNAs have been found to play a crucial role in development, differentiation, apoptosis, and metabolism and are involved in the pathogenesis of many human diseases. 2,3 Bioinformatics studies suggest that miRNAs may regulate Ͼ60% of all human genes. 4,5 Studies have shown that overexpression of miRNA lin-4 increases longevity in Caenorhabditis elegans, whereas loss of lin-4 leads to a

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“…Selective culture conditions (68,70) and lectins or antibodies attached to magnetic beads (20,22,43,44,51,74,106) have proven useful in this regard. However, incomplete specificity for endothelial cells and removal of cell surface epitopes during dissociation are limitations of this approach.…”

mentioning

confidence: 99%

Expression of genes involved in vascular development and angiogenesis in endothelial cells of adult lung

Favre

Mancuso

Maas

et al. 2003

American Journal of Physiology-Heart and Circulatory Physiology

123

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Profiling gene expression in endothelial cells advances the understanding of normal vascular physiology and disease processes involving angiogenesis. However, endothelial cell purification has been challenging because of the difficulty of isolating cells and their low abundance. Here we examine gene expression in endothelial cells freshly isolated from lung capillaries after in vivo labeling with fluorescent cationic liposomes and purification by fluorescence-activated cell sorting (FACS). Of the 39,000 genes and expressed sequence tags evaluated on custom oligonucleotide arrays, 555 were enriched in endothelial cell fraction. These included familiar endothelial cell-associated genes such as VEGF, VEGF receptor (VEGFR)-1, VEGFR-2, angiopoietin-2, Tie1, Tie2, Edg1 receptor, VE-cadherin, claudin 5, connexin37, CD31, and CD34. Also enriched were genes in semaphorin/neuropilin (Sema3c and Nrp1), ephrin/Eph (ephrin A1, B1, B2, and EphB4), delta/notch (Hey1, Jagged 2, Notch 1, Notch 4, Numb, and Siah1b), and Wingless (Frizzled-4 and Tle1) signaling pathways involved in vascular development and angiogenesis. Expression of representative genes in alveolar capillary endothelial cells was verified by immunohistochemistry. Such expression reflects features that endothelial cells of normal lung capillaries have in common with embryonic and growing blood vessels. About half of the enriched genes, including exostosin 2, lipocalin 7, phospholipid scramblase 2, pleckstrin 2, protocadherin 1, Ryk, scube 1, serpinh1, SNF-related kinase, and several tetraspanins, had little or no previous association with endothelial cells. This approach can readily be used to profile genes expressed in blood vessels in tumors, chronic inflammation, and other sites in which endothelial cells avidly take up cationic liposomes.

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Isolation of Endothelial Cells from Brain, Lung, and Kidney: Expression of the Multidrug Resistance P-Glycoprotein Isoforms

Cited by 99 publications

References 38 publications

Side population cells and Bcrp1 expression in lung

Side population cells and Bcrp1 expression in lung

miR-335 and miR-34a Promote Renal Senescence by Suppressing Mitochondrial Antioxidative Enzymes

Expression of genes involved in vascular development and angiogenesis in endothelial cells of adult lung

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