Variation of UCB CD34+ in erythroid differentiation potential
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ABSTRACT:
PROVIDER: PRJNA763126 | ENA |
REPOSITORIES: ENA
Dataset's files
| Action | DRS | |||
|---|---|---|---|---|
| SRR15886461_1.fastq.gz | Fastqsanger.gz | |||
| SRR15886461_2.fastq.gz | Fastqsanger.gz | |||
| SRR15886462_1.fastq.gz | Fastqsanger.gz | |||
| SRR15886462_2.fastq.gz | Fastqsanger.gz | |||
| SRR15886463_1.fastq.gz | Fastqsanger.gz |
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Project description:Ex vivo manufactured red blood cells (RBC) generated from immortalized erythroid cell lines which can semi-infinitely grow are expected to become a significant alternative in future transfusion therapies. To establish those cell lines, ectopic expression of human papilloma virus (HPV) E6/E7 gene has successfully been employed. In order to induce differentiation and maturation of the immortalized cell lines, terminating the HPV-E6/E7 expression through a gene induction system has been believed to be essential. Here we report that erythroid cell lines established from human bone marrow using simple overexpression of HPV-E6/E7 is capable of normal erythroid differentiation without turning the gene expression off. Newly established cell lines, Erythroid Lines from Lund University (ELLU), are able to differentiate towards mature cells including enucleated reticulocytes upon changing the culture condition but without terminating HPV-E6/E7 gene expression. ELLU cells are heterogeneous, and unexpectedly, clones expressing adult hemoglobin rapidly differentiate but produced cells are fragile while other clones with fetal hemoglobin start expressing adult hemoglobin upon differentiation and give rise to more mature cells. Our findings propose an alternative method to establish immortalized human erythroid cell lines and describe novel cellular characteristics for desired functionally competent clones.
2022-01-31 | GSE184083 | GEO
Project description:We investigated the miRNA expression in ex vivo human erythroid cultures from umbilical cord blood (UCB)-derived CD34 cells. Hypothetically, the decline of certain miRNAs may promote erythropoiesis by unblocking expression of key functional proteins while the up-regulation of other miRNAs may block commitment to non-erythroid lineages. Therefore, discovering the patterns and sequence of miRNA expression during hematopoietic differentiation will provide insights about the functional roles of these tiny noncoding genes. Keywords: miRNA expression profiling
2007-04-01 | GSE6000 | GEO
Project description:Preeclampsia (PE) has been associated with placental dysfunction, resulting in foetal hypoxia, accelerated erythropoiesis and increased erythroblast count in the umbilical cord blood (UCB). Although the detailed effects remain unknown, placental dysfunction can also cause inflammation, nutritional and oxidative stress in the fetus that can affect erythropoiesis. Here, we compared the expression of surface adhesion molecules and erythroid differentiation capacity of UCB hematopoietic stem/ progenitor cells (HSPCs), UCB erythroid profiles along with transcriptome and proteome of these cells between male and female foetuses from PE and normotensive pregnancies. While no significant differences were observed in UCB HSPC migration/ homing and in vitro erythroid colony differentiation, the UCB HSPC transcriptome and the proteomic profile of the in vitro differentiated erythroid cells differed between PE vs normotensive samples. Accordingly, despite absence of significant differences in the UCB erythroid populations in male or female foetuses from PE or normotensive pregnancies, transcriptional changes were observed during erythropoiesis, particularly affecting male foetuses. Pathway analysis suggested deregulation in mTORC1/AMPK signaling pathways controlling cell cycle, differentiation and protein synthesis. These results associate PE with transcriptional and proteomic changes in foetal HSPCs and erythroid cells that may underlie the higher erythroblast count in the UCB in PE.
2019-06-20 | PXD010364 | Pride
Project description:Immunoregulatory potential of erythroid nucleated cells induced from CD34+ progenitors from bone marrow
| PRJNA967257 | ENA
Project description:Hematopoietic stem and progenitor cells are a rare, self-renewing bone marrow resident population capable of giving rise to all circulating hematopoietic cells. They can be used therapuetically for reconstituting defective or ablated hematopoietic systems following chemotherapy, and for inducing tolerance toward allografts of the same haplotype as the HSC donor. There are several sources for HSCs, such as the adult bone marrow, or umblical cord blood, which is more replete with such HSCs. However, HSCs obtained from such sources may be immunogenic, especially if isolated from adult bone marrow. To overcome this issue, our lab has establsihed human induced pluripotent stem cell-derived HPCs with the hope of creating a nonimmunogenic, readily available and unlimited source of HSCs to use for therapy. The goal of this study was to compare the gene expression profiles of naturally found HSCs (UCB-CD34+ HSCs) and HPCs differentiated from 4 different human iPS cell lines (iPS-HPCs), so as to determine the variation between the four iPS-HPCs and whether there were any differences between these HPCs and naturally found HSCs. We utilized 4 iPS cells for this study (detailed descriptions are provided below). iPS cells were differentiated into hematopoietic progenitor cells by coculture on OP9 stromal cells, followed by enrichment of CD34+ cells through immunomagnetic bead separation. The UCB-CD34+ cells were isolated from frozen cord samples through immunomagnetic bead separation. Total RNA was isolated and human gene Affymetrix ST 1.0 arrays performed at the University of Iowa DNA core facility. Data was analyzed, normalized and plotted on BRB Array Tools.
2013-10-23 | E-GEOD-46888 | biostudies-arrayexpress
Project description:Fine-resolution differentiation trajectories of adult human hematopoietic stem cells (HSC) involved in the generation of red cells is critical for understanding dynamic developmental changes that accompany human erythropoiesis. Using Single-cell RNA sequencing (scRNA-seq) of primary human terminal erythroid cells (CD34−CD235a+) isolated directly from adult bone marrow (BM) and umbilical cord blood (UCB), we documented the transcriptome of terminally differentiated human erythroblasts at unprecedented resolution. The insights enabled us to distinguish polychromatic erythroblasts (PolyE) at the early and late stages of development as well as the different development stages of orthochromatic erythroblasts (OrthoE). We further identified a set of putative regulators of terminal erythroid differentiation and functionally validated three of the identified genes, AKAP8L, TERF2IP and RNF10, by monitoring cell differentiation and apoptosis. We documented that knockdown of AKAP8L suppressed the commitment of HSC to erythroid lineage and cell proliferation, and delayed differentiation of colony forming unit-erythroid (CFU-E) to the proerythroblast stage (ProE). In contrast, the knockdown of TERF2IP and RNF10 delayed differentiation of PolyE to OrthoE stage. Taken together, the convergence and divergence of the transcriptional continuums at single-cell resolution underscore the transcriptional regulatory networks that underlie human fetal and adult terminal erythroid differentiation.
2020-05-25 | GSE150774 | GEO
Project description:The associated publication describes a RUNX3-dependent defect in primary human CD34+ HSPC differentiation into the erythroid lineage. This study was designed to determine which genes are potential targets of RUNX3, and could contribute to regulation of erythropoiesis.
2019-03-01 | GSE119264 | GEO
Project description:The characteristics of global gene expression patterns during umbilical cord blood (UCB)-CD34+ stem cell-derived erythropoiesis are not clearly elucidated. In this study, UCB-stem cells were grown in liquid culture as a model for this process. We observed a high proliferative capacity of UCB-stem cells producing over 10 billion viable cells in culture. Normal erythroid maturation was confirmed by increased expression of CD71 and CD235a biomarkers. Furthermore, the gamma- to beta-globin gene switch was observed around day 45 indicating extended gamma-globin gene expression in fetal erythroid cells. To study global gene expression patterns, microarray analysis was performed on days 21, 42, 49 and 56. Forty-five transcription factors were silenced during the culture period (Profile-1) including CUX1 and HES5 among others. Conversely, 30 transcription factors were activated by day 56 (Profile-2) such as KLF1, GATA1, and MAFB. Both datasets were analyzed further using the MIMI Cytoscape platform which defined transcription factor networks around KLF4 and GATA2 from Profile-1 genes and KLF1 and GATA1 for Profile-2 genes. The characteristics of UCB-CD34+ stem cells including high proliferative capacity and prolonged gamma-globin expression combined with novel transcription factor networks suggest novel mechanisms of fetal UCB-stem cell-derived erythropoiesis.
2013-08-02 | GSE49438 | GEO
Project description:CD34-positive cells from peripheral blood were culture for 5 days in erythroid differentiating medium. Four progenitor stages were sorted by FACS using the established cell surface markers CD34 and CD36 with CD117, CD71, and CD105 (Yan H, Am J Hematol, 2021). A comprehensive analysis of the proteome of these four erythroid progenitor stages was done in quadruplicate using a label free proteomic approach.
2023-10-30 | PXD042615 | Pride
Project description:The characteristics of global gene expression patterns during umbilical cord blood (UCB)-CD34+ stem cell-derived erythropoiesis are not clearly elucidated. In this study, UCB-stem cells were grown in liquid culture as a model for this process. We observed a high proliferative capacity of UCB-stem cells producing over 10 billion viable cells in culture. Normal erythroid maturation was confirmed by increased expression of CD71 and CD235a biomarkers. Furthermore, the gamma- to beta-globin gene switch was observed around day 45 indicating extended gamma-globin gene expression in fetal erythroid cells. To study global gene expression patterns, microarray analysis was performed on days 21, 42, 49 and 56. Forty-five transcription factors were silenced during the culture period (Profile-1) including CUX1 and HES5 among others. Conversely, 30 transcription factors were activated by day 56 (Profile-2) such as KLF1, GATA1, and MAFB. Both datasets were analyzed further using the MIMI Cytoscape platform which defined transcription factor networks around KLF4 and GATA2 from Profile-1 genes and KLF1 and GATA1 for Profile-2 genes. The characteristics of UCB-CD34+ stem cells including high proliferative capacity and prolonged gamma-globin expression combined with novel transcription factor networks suggest novel mechanisms of fetal UCB-stem cell-derived erythropoiesis. Cd34 cell from umbilical cord blood were grown in three independent cultures using the one-phase protocol. Cells were cultured in aMEM containing 30% fetal bovine serum, 1% deionized BSA with penicillin (100 U/mL) and streptomycin (0.1 mg/mL) at 37°C and 5% CO2. The following growth factors were added on day 0: stem cell factor (50 ng/mL), interleukin-3 (10 ng/mL) and erythropoietin (4 U/mL). Approximately 1.5 million cells were harvested from each culture (triplicate samples) on day 21, 42, 49 and 56 for microarray analysis on the whole-genome Illumina HumanWG-12 V4 Expression BeadChip.
2013-08-02 | E-GEOD-49438 | biostudies-arrayexpress