Project description:Expression profiling of lung derived mesenchymal stromal cells to lung fibrblasts and cord blood derived mesenchymal stromal cells We have previously isolated mesenchymal stromal cells (MSCs) from the tracheal aspirates of premature neonates with respiratory distress. While isolation of MSCs correlates with the development of bronchopulmonary dysplasia, the physiologic role of these cells remains unclear. To address this, we further characterized the cells, focusing on the issues of gene expression, origin and cytokine expression. Microarray comparison of early passage neonatal lung MSC gene expression to cord blood MSCs and human fetal and neonatal lung fibroblast lines demonstrated that the neonatal lung MSCs differentially expressed 971 gene probes compared to cord blood MSCs, including the transcription factors Tbx2, Tbx3, Wnt5a, FoxF1 and Gli2, each of which have been associated with lung development. Compared to lung fibroblasts, 710 gene probe transcripts were differentially expressed by the lung MSCs, including IL-6 and IL-8/CXCL8. Further, neonatal lung MSCs exhibited a pattern of Hox gene expression distinct from cord-blood MSCs but similar to human fetal lung fibroblasts, consistent with a lung origin. Together, these data suggest that MSCs isolated from neonatal tracheal aspirates originate in the lung and are distinct from lung fibroblasts.
Project description:Bronchopulmonary dysplasia remains one of the most common complication of prematurity, despite significant improvements in perinatal care. Functional modeling of human lung development and disease, like BPD, is limited by our ability to access the lung and to maintain relevant stem cell populations in culture. Single cell RNA-sequencing confirmed the presence of epithelial cells in tracheal aspirates obtained from intubated neonates. Using combined SMAD signaling inhibition and mTOR inhibition neonatal tracheal-aspirate derived (nTAD) basal stem cells can be expanded long-term and retain the ability to differentiate into pseudo-stratified airway epithelium. Conclusions: Our data demonstrate that neonatal tracheal aspirate-derived epithelial cells can provide a novel ex vivo human cellular model to study neonatal lung development and disease.
Project description:Bronchopulmonary dysplasia remains one of the most common complication of prematurity, despite significant improvements in perinatal care. Functional modeling of human lung development and disease, like BPD, is limited by our ability to access the lung and to maintain relevant stem cell populations in culture. Single cell RNA-sequencing confirmed the presence of epithelial cells in tracheal aspirates obtained from intubated neonates. Using combined SMAD signaling inhibition and mTOR inhibition neonatal tracheal-aspirate derived (nTAD) basal stem cells can be expanded long-term and retain the ability to differentiate into pseudo-stratified airway epithelium. Conclusions: Our data demonstrate that neonatal tracheal aspirate-derived epithelial cells can provide a novel ex vivo human cellular model to study neonatal lung development and disease.
Project description:We compared differential gene expression in tracheal aspirates collected mechanically ventilated subjects with COVID-19 ARDS to gene expression in tracheal aspirates from: 1) subjects with ARDS from other casues and 2) mechanically ventilated controls without evidence of pulmonary disease.
Project description:The purpose of this experiment was to determine the transcriptional differences between neural progenitor cells from neonatal lung injury mice vs. control mice, as well as neonatal lung injury mice treated with umbilical-cord mesenchymal stromal cell extracellular vesicles vs. neonatal lung injury mice treated with placebo (PBS). Neural progenitor cells were isolated from the subventricular zone and hippocampus and cultured for 2 consecutive neurosphere assays. RNA was then extracted from the cells, and the microarray labelling, hybridization, and scanning was conducted by the Génome Québec Innovation Centre (Montréal, Canada).
Project description:Neonatal Tracheal Aspirate-derived Airway Epithelial Cells (nTAECs) were isolated utilizing fresh tracheal suction aspirates from preterm (born at 23-28wks gestational age) and term infants (born at 37-42wks gestation). Included infants (especially term infants) did not have significant lung disease at the time of collection. Isolated cells were plated in 804g-cell-derived matrix coated T25, then expanded to a T75 flask and subsequently cultured in transwell inserts (0.4μm size pore) at 105 live cells/insert under air-liquid interface (ALI). Basal nTAECs have the capacity to differentiate into the full repertoire of mature airway epithelial lineages (ciliated, secretory club and mucus secreting goblet cells) during ALI culture. Cells were exposed to 60% O2 or room air control (21% O2) for 7 days (ALI day 7 to 14). On ALI day 14, cells will be harvested from the transwells and prepared for proteomic analysis using Data-independent acquisition (DIA) proteomics analysis. The samples will undergo several processing steps including chloroform/methanol extraction with trypsin digestion, Orbitrap Exploris 480 mass spectrometry analysis and eventual library construction and bioinformatics analysis. Utilizing 5 donor samples for each group, the proteomic workflow compared 4 exposure groups: a) Term nTAECs in Normoxia, b) Term nTAECs Hyperoxia, c) Preterm nTAECs in Normoxia, d) Preterm nTAECs in Hyperoxia.
Project description:Extreme preterm infants are a growing population in the neonatal intensive care unit. Multiple factors play a role in preterm birth, resulting in complications including severe bronchopulmonary dysplasia (sBPD) without or with and pulmonary hypertension (BPD-PH). The goal of this study was to identify biomarker signatures associated with sBPD and BPD-PH. We analyzed profiles in tracheal aspirates (TAs) from 46 extremely preterm infants receiving invasive mechanical ventilation (25 sBPD, 21 BPD-PH) . We found specific miRNA signatures in TAs that may serve as biomarkers for the two disease phenotypes.
