ABSTRACT: Comparative gene expression analyses of purified CD14+ monocytes from human umbilical cord blood, CD14+ monocytes from adult peripheral blood and the cell therapy product DUOC-01
Project description:DUOC-01 is a macrophage-like cell therapy product manufactured from human umbilical cord blood (CB) mononuclear cells (MNCs) and intended for use in treatment of demyelinating diseases. During the 21-day manufacturing process, most initiating MNCs die in culture, but a highly active, adherent population of cells that resemble hematopoietic lineage macrophage in morphology, phagocytic activity, and antigen expression emerges; this population comprises the final DUOC-01 product [Kurtzberg et al. 2015]. Using the cuprizone-mediated murine demyelination model, we have found that DUOC-01 cells are capable of accelerating the remyelination process. In the same demyelination model, human cord blood-derived CD14+ cells (CBCD14) were less active. In another cellular therapy project, we have found that CBCD14 protects brain cells from oxygen-glucose-deprivation (OGD), but adult peripheral blood-derived CD14+ monocytes (PBCD14) did not. To address the functional differences between these cell types, we have analyzed differential gene expression by these three cell types. We used four cord blood (CB) samples (from Carolinas Cord Blood Bank) that were collected within 24hrs of delivery. Then, highly purified CB CD14+ monocytes were isolated by flow sorting. We collected the peripheral blood (PB) from four healthy volunteers and isolated the purified PB CD14+ monocytes by cell sorting. Three separate batches of cultured DUOC-01 cells were collected from Robertson Cell and Translational Cell Therapy cGMP facility. Total RNAs were isolated at the same time and kept frozen. RNA samples were given to Duke Microarray core facility to analyse the gene expression profile using Affymetrix GeneChip® Human Genome U133 Plus 2.0 Assay.
Project description:DUOC-01 is a macrophage-like cell therapy product manufactured from human umbilical cord blood (CB) mononuclear cells (MNCs) and intended for use in treatment of demyelinating diseases. During the 21-day manufacturing process, most initiating MNCs die in culture, but a highly active, adherent population of cells that resemble hematopoietic lineage macrophage in morphology, phagocytic activity, and antigen expression emerges; this population comprises the final DUOC-01 product [Kurtzberg et al. 2015]. Using the cuprizone-mediated murine demyelination model, we have found that DUOC-01 cells are capable of accelerating the remyelination process. In the same demyelination model, human cord blood-derived CD14+ cells (CBCD14) were less active. In another cellular therapy project, we have found that CBCD14 protects brain cells from oxygen-glucose-deprivation (OGD), but adult peripheral blood-derived CD14+ monocytes (PBCD14) did not. To address the functional differences between these cell types, we have analyzed differential gene expression by these three cell types.
Project description:We sought to determine the impact of chorioamnionitis exposure on term neonatal monocyte transcription. RNA-seq was performed on term healthy and chorioamnionitis-exposed umbilical cord blood purified CD14+ monocytes under unstimulated and LPS stimulated conditions.
Project description:A phenotypically and functinoally distinct subset of human blood dendritic cells expressing CD11b is specific of the neonatal environment. We have employed whole genome microarray expression profiling to identify the specific gene signature of CD11b+ cord blood dendritic cells as compared to their adult peripheral blood counterparts. Peripheral blood adult cDC2 (CD20- CD11c+ CD14- BDCA1+ CD11b- ), neonatal cord blood cDC2 (CD20- CD11c+ CD14- BDCA1+ CD11b-) and neonatal cord blood cDC2b (CD20- CD11c+ CD14- BDCA1+ CD11b+) were FACS purified from BDCA1+ magnetically. Neonatal monocytes (CD11c+ CD14+) and neonatal naive T cells (CD3+ CD4+ CD56- CD25- CD45RO-) were used as controls.
Project description:We sought to determine the impact of chorioamnionitis exposure on the neonatal monocyte H3K4me3 histone modification landscape over the course of fetal and neonatal immune system development using ChIP-seq. H3K4me3 ChIP-seq was performed on umbilical cord blood purified CD14+ monocytes from healthy and chorioamnionitis-exposed extremely preterm neonates (under 30 weeks gestation), late preterm neonates (30-36 weeks gestation), and term neonates (37+ weeks gestation).
Project description:To identify the differences between human umbilical cord blood and peripheral blood monocytes, we performed unsupervised bioinformatic analyses by microarrays.
Project description:We isolated resting platelets from full term umbilical cord blood and adult peripheral blood. We used DIA mass spectrometry to analyze the whole proteome and phosphoproteome from this platelet samples.
Project description:Here, we performed proteomic profiling of the maternal peripheral blood and umbilical cord blood from 6 pregnant women positive for SARS-CoV-2 and 6 pregnant women negative for SARS-CoV-2, and examined viral RNA and DNA copies of SARS-CoV-2 sequences in the umbilical cord and placental tissues.
