Transcription profiling of human umbilical cord blood TO study expression OF background, progenitor AND stem cell populations
ABSTRACT: Gene expression study of populations of primary human hematopoietic cells from human cord blood including background, progenitor and stem cells. Experiment Overall Design: This experiment include 4 samples each with 3 replicates on 2 platforms for a total of 24 GEO Samples
Project description:Gene expression study of populations of primary human hematopoietic cells from human cord blood including background, progenitor and stem cells. Keywords: other Overall design: This experiment include 4 samples each with 3 replicates on 2 platforms for a total of 24 GEO Samples
Project description:There were no studies about gene expression of umbilical cord tissue before. We performed this study to identify the gene expression of umbilical cord tissue. Overall design: Umbilical cord were obtained from three newborns. Gene expression profiles were conducted using cDNA microarray analysis.
Project description:Here we compare the gene expression profiles of two distict, fluorescently identified neuronal populations, the motor neruons (MN) and the decending commissural interneurons (dCIN) of the neonatal rat spinal cord isolated with laser microdissection. These two populations participate in the neuronal networks in the spinal cord where they have destinct functions in shaping (dCINs) and transferring the output to the muscles (MNs) during locomotion. We wished to determine how the functinal differences in these nerurons are manifested at the gene expression level. Experiment Overall Design: Neurons were retrogradely labelled with rhodamine through application of the tracer to cut axons followed by incubation in oxygenated Ringer solution. Spinal cord were then snap frozen, cryosectioned and fluorescent single cells were laser dissected into a collector tube to a total of ca. 50-200 cells pr sample. RNA was then extracted, two round amplified and hybridized to Affymatrix RNU-34 chips. 22 samples were hybridized onto chips, 7 MNs, 7 CINS and 8 MIX.
Project description:Maternal smoking doubles the risk of delivering a low birth weight infant. The purpose of this study was to analyze differential gene expression in umbilical cord tissue as a function of maternal smoking, with an emphasis on growth-related genes. We recruited 15 pregnant smokers and 15 women who never smoked during pregnancy to participate RNA was isolated from umbilical cord tissue collected and snap frozen at the time of delivery. Microarray analysis was performed using the Affymetrix GeneChip Scanner 3000.Six hundred seventy-eight probes corresponding to 545 genes were differentially expressed (i.e., an intensity ratio that exceeded +/-1.3 and a corrected significance value p < 0.005) in tissue obtained from smokers versus nonsmokers. Genes important for fetal growth, angiogenesis, or development of connective tissue matrix were up-regulated among smokers. The most highly up-regulated gene was CSH1, a somatomammotropin gene. Two other somatomammotropin genes (CSH2 and CSH-L1) were also up-regulated. The most highly down-regulated gene was APOBEC3A; other down-regulated genes included those that may be important in immune and barrier protection. PCR validation of the three somatomammotropin genes showed a high correlation between qPCR and microarray expression. Consequently, maternal smoking may be associated with altered gene expression in the offspring. Experiment Overall Design: 30 samples of two groups consiting of 15 smokers (smk) and 15 nonsmokers (nonsmk) were analyzed without replicates. Experiment Overall Design: Note: Non-corrupt .CEL files are not available for the Samples in this study.
Project description:A non-controversial and non-invasive source of adult stem cells (ASCs), particularly hematopoietic stem cells (HSCs) and mesenchymal stem cells (MSCs) is human umbilical cord blood. HSCs derived from cord blood have been used for treating leukemia and other blood disorders for the last 30 years. While the presence of MSCs in cord blood is limited, umbilical cord has been found to be promising source of MSCs. However, the cord is an anatomically complex organ and potential isolation of MSCs from its various parts has not been fully explored. In this study we dissected the cord into cord placenta junction (CPJ), cord tissue (CT), and Wharton’s jelly (WJ) and isolated stem cells. These cells exhibited fibroid morphology, expressed MSC-specific markers including CD90, CD73, CD105, CD44, and CD29 and differentiated into chondrogenic, osteogenic, myogenic and neurogenic lineages. In addition, they all expressed pluripotency genes, OCT4, Nanog, Sox2 and KLF4 but expression of these markers was highest in CPJ followed by WJ and CT. CPJ-MSCs also had higher rate of proliferation compared to WJ- and CT-MSCs. Proliferation of WJ- and CT-MSCs was markedly decreased upon passaging with concomitant decrease in expression of MSC and pluripotency markers. Based on their greater self-renewal potential, CPJ-MSCs could be superior to WJ- and CT-MSCs for the applications in therapeutic and regenerative medicine. Overall design: Human umbilical cord derived MSCs were cultured in vitro. Gene expression of cells originating from each of cord-placenta junction and Wharton's jelly were compared to cells originating from cord tissue.
Project description:AML1-ETO expression in normal human umbilical cord blood CD34+ cells leads to long-term proliferation of an early self-renewing primitive progenitor cell with multilineage potential and stem cell ability, but these cells do not induce leukemia in immunodeficient mice. This comparative microarray study was initiated to determine the differences in the transcriptome of AML-ETO-expressing CD34+ cells after extended culture in vitro, using normal cord blood cells expanded for 6-8 weeks in vitro and subsequently purified for the CD34+ population as the control comparison. Experiment Overall Design: We have established a culture system whereby we retrovirally transduce human CD34+ cells, obtained from cord blood, with the leukemia fusion gene AML1-ETO. Cells expressing this fusion protein are able to proliferate long-term in vitro in a cytokine dependent manner. AML1-ETO-expressing cord blood cells have a large population of primitive self-renewing CD34+ cells with continued abnormal differentiation. We grow these cells in serum-free conditions using the BIT supplement from Stem Cell Technologies. For the current experiments we used cell cultures that had been proliferating in vitro for 8-12 weeks, in a cytokine cocktail of SCF, TPO, FLT3L, IL-6 all at 20 ng/mL and IL-3 at 10 ng/mL. Control cord blood samples that were CD34 purified were expanded for 5-8 weeks in the same culture media as used for AML1-ETO cells. All samples were magnetically selected for the CD34+ population, returned to culture, and one week later again selected for CD34+ cells and then lysed for RNA isolation.
Project description:Analysis of umbilical cord tissue in newborns of type 1 diabetic mothers at gene expression level. The hypothesis tested in the present study was that intrauterine diabetic milieu may effect of fetal umbilical cord gene expression, and via umbilical cord, the alterations may be produced in other fetal tissues as well. Results provide an information of the differentially expressed genes and enriched pathways, such as the dowregulated genes on the pathway on blood vessel development in umbilical cords from diabetic pregnancies. Umbilical cord tissue was collected after elective ceasarean section and was immediately flash frozen in liquid nitrogen and stored at -80C until total RNA extraction from the whole tissue sample. Six cords exposed to maternal diabetes (DM) and six control cords (C) from healthy pregnancies were analyzed.
Project description:Goals/objectives: to identify various gene expression in B cell subsets derived from human PBMC and cord blood Sample: human PBMC and human cord blood Cell subsets: memory B cell (CD20+CD10-CD27+), naive B cell (CD20+CD10-CD27-), CD21lo transitional B cell (CD20+CD10+CD27-CD21lo), CD21hi transitional B cell (CD20+CD10+CD27-CD21hi) Number of sample: PBMC (pooled from 2 donors), cord blood (pooled from 2-3 donors) Number of repeats: 2 Microarray: Affymetrix HU133Plus2 RNA extraction: Trizol (Invitrogen) RNA preparation/hybridization: All according to affymetrix protocol
Project description:Human umbilical cord blood cells (UCB) is an important alternative resource for the hematopoietic stem cells in treatment of leukemia and other non-malignant diseases. The failure of hematopoiesis reconstruction by the UCB is known to be associated with several clinicopathological features of host patients. There are very few reports available, however, seeking for the association with the quality of umbilical cord blood cells (UCB) themselves. Here we try to address the quality of UCBs by transfusion to the lethally irradiated immunocompromized mice. The cryopreserved CD34+ UCBs cells from twelve different single human donors were transplanted to the irradiated NOD/shi-scid Jic mice. In parallel, total RNAs of the UCBs were subjected to the gene expression profiling with oligonucleotide microarrays. The UCBs from three donors failed to establish the engraftment in the host mice whereas other 9 UCBs succeeded to various extents. Oligonucleotide microarray analysis indicated that 71 genes, including HOXB4 and ETS2, were specifically overexpressed and 23 genes were suppressed more than 2 fold in the successful UCBs comparing with the failed ones (p<0.005, Student’s t-test). Functional annotation analyses revealed that the genes mediated cell growth and cell cycle regulations were enriched in successful UCBs comparing with the failed ones. Our results suggest that the hematopoiesis reconstitution ability may vary among the cryopreserved UCBs and the quality can be distinguished with certain sets of gene expression. Overall design: The gene expression in human umbilical cord blood cells (UCBs) was measured before transplantation to the lethally irradiated immunocompromized mice. UCBs from 12 individuals were examined.
Project description:screening of signature deterimes the individual variations in the therapeutic efficacy of human umbilical cord blood-derived mesenchymal stem cells There is paucity of information whether human umbilical cord blood-derived mesenchymal stem cells (hUCB-MSCs) from separate donors might have different effects on improving myocardial repair after myocardial infarction (MI). We screened cell surface genes by the comparing the cells that showed the best and worst efficacy, respectively, in repairing the infarcted myocardium of rats.