Unknown,Transcriptomics,Genomics,Proteomics

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Transcription profiling of rat young and aged mesenchymal stem cells response to dexamethasone

ABSTRACT: To explore the molecular events that underline skeletal physiology during aging we catalogued the profile of gene expression in ex vivo cultured MSCs derived from 3 and 15 month old rats and challenged with dexamethasone (Dex). RNA retrieved from these cells was analyzed using Affymetrix Gene Chips to compare the effect of Dex on gene expression. Experiment Overall Design: Mesenchymal Stem Cells were collected in the following manner: the bone marrow cells were flushed from the long bones, collected and prepared for single cells suspension. Cell pooled from six rats in each group were cultured in 75 cm2 flasks (Falcon, USA) containing 20 ml of growth medium, Dulbeccoâs Modified Essential Medium (DMEM) containing 10% heat-inactivated fetal calf serum (FCS). Under these conditions, the hematopoietic cells died and the cultures finally remained only with cells forming the adherent stromal fibroblast-like layer. Cells were plated and after one week of culturing were trypsinized, single cell suspensions were re-cultured for 7 days and were grown in absence or presence of 10-8 M Dexamethasone (Dex). On day 14 cells were harvested for RNA extraction. Experiment Overall Design: The groups included: young (3 month) untreated females, young (3 month) untreated males, young treated (Dex) females, old (15 month) untreated rats, old treated (Dex) rats. Experiment Overall Design: Total RNAs were extracted from the four experimental groups using the TRIzol and cRNA prepared according to Affymetrix protocols. Gene expression was measured by hybridization to Affymetrix RAE230A Gene Chip DNA microarrays .

ORGANISM(S): Rattus norvegicus

SUBMITTER: Dafna Benayahu

PROVIDER: E-GEOD-3339 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Transcriptional profiling of mesenchymal stromal cells from young and old rats in response to Dexamethasone.

Akavia Uri David UD Shur Irena I Rechavi Gideon G Benayahu Dafna D

BMC genomics 20060427

<h4>Background</h4>Marrow-derived stromal cells (MSCs) maintain the capability of self-renewal and differentiation into multiple lineages in adult life. Age-related changes are recognized by a decline in the stemness potential that result in reduced regeneration potential of the skeleton. To explore the molecular events that underline skeletal physiology during aging we catalogued the profile of gene expression in ex vivo cultured MSCs derived from 3 and 15 month old rats. The ex vivo cultured c ...[more]

PMID: 16643645

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Project description:Introduction: Renal ischemia-reperfusion (IR) causes acute kidney injury (AKI) with high mortality and morbidity. The objective of this study was to ameliorate kidney IR injury and identify novel biomarkers for kidney injury and repair. Methods: Left renal ischemia was induced in rats by clamping renal artery for 45 minutes, followed by reperfusion and right nephrectomy. Thirty minutes prior to ischemia, rats (n=8/group) received Valproic Acid (150 mg/kg; VPA), Dexamethasone (3 mg/kg; Dex) or Vehicle (Saline) intraperitoneally. Animals were sacrificed at 3h, 24h or 120h post- IR and blood, urine and kidney were collected. Results: Serum creatinine (mg/dL) at 24 h IR in VPA (2.7±1.8) and Dex (2.3±1.2) was reduced (P<0.05) compared to Vehicle (3.8±0.5). At 3h post-IR, urine albumin (mg/ml) was higher in Vehicle (1.47±0.10), VPA (0.84±0.62) and Dex (1.04±0.73) compared to uninjured/untreated control (0.14±0.26) group. At 24h post-IR urine Lipocalin-2 (µg/ml) was significantly higher (P<0.05) in VPA, Dex and Vehicle groups (9.61-11.36) compared to uninjured/untreated control (0.67±o.29); also, Kidney Injury Molecule-1 (KIM-1; ng/ml) was significantly higher in VPA, Dex and Vehicle groups (13.7-18.7) compared uninjured/untreated control (1.7±1.9). KIM-1 levels were significantly (P<0.05) higher in all groups compared to uninjured/untreated control levels. Histopathology at 3h post IR demonstrated (P<0.05) reduction in ischemic injury in the renal cortex in VPA (Grade 1.6± 1.5) compared to Vehicle (Grade 2.9±1.1) group. Inflammatory cytokines IL1β and IL6 were down-regulated in VPA and Dex groups. BCL2 was higher in VPA group. DNA microarray analysis demonstrated reduced stress response and injury, and improved recovery related gene expression in the kidneys of VPA treated animals. Conclusions: VPA administration reduced kidney IR injury and improved regeneration. KIM-1 and Lipocalin-2 appear to be promising early urine biomarkers of acute ischemic kidney injury.

Dataset Information

Transcription profiling of rat young and aged mesenchymal stem cells response to dexamethasone

Publications

Transcriptional profiling of mesenchymal stromal cells from young and old rats in response to Dexamethasone.

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