scholarly journals Reduced angiovasculogenic and increased inflammatory profiles of cord blood cells in severe but not mild preeclampsia

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Seonggeon Cho ◽  
Young-Doug Sohn ◽  
Sangsung Kim ◽  
Augustine Rajakumar ◽  
Martina L. Badell ◽  
...  

AbstractPreeclampsia (PE) is a prevalent pregnancy disorder that leads to high maternal and fetal morbidity and mortality. While defective vascular development and angiogenesis in placenta are known as crucial pathological findings, its pathophysiological mechanism remains elusive. To better understand the effects of PE on angio-vasculogenesis and inflammatory networks in the fetus and to identify their biological signatures, we investigated the quantitative and functional characteristics of cord blood-derived mononuclear cells (CB-MNCs) and CD31-positive MNCs. Flow cytometry analysis demonstrated that the CB-MNCs from the severe PE group had significantly decreased number of cells expressing CD3, CD11b, CD14, CD19, KDR, and CD31 compared with the normal group. Quantitative real time PCR (qRT-PCR) shows down-regulation of the major angiogenic factor VEGFA in MNCs and CD31+ MNCs in severe PE. The major inflammatory cytokines IL1 was highly upregulated in CD31+ CB-MNCs in the severe PE patients. Mild PE patients, however, did not display any significant difference in expression of all measured angiogenic genes and most inflammatory genes. These findings show distinct angiogenic and inflammatory signatures from severe PE, and they may play a significant role in the pathogenesis of vascular defects in placenta of severe PE.

2020 ◽  
Vol 14 ◽  
pp. 117793222091330
Author(s):  
LM Avila-Portillo ◽  
F Aristizabal ◽  
S Perdomo ◽  
A Riveros ◽  
B Ospino ◽  
...  

Biosimilars of granulocyte colony-stimulating factor (G-CSF) have been routinely introduced into clinical practice. However, not functional genomics characterization has been performed yet in comparison with the innovator G-CSF. This study aimed to evaluate the transcriptomic changes in an in vitro model of umbilical cord blood cells (UBC) exposed to G-CSF for the identification of their modulated pathways. Umbilical cord blood cells–derived mononuclear cells (MNCs) were treated with biosimilar and innovator G-CSF for further gene expression profiling analysis using a microarray-based platform. Comparative analysis of biosimilar and innovator G-CSF gene expression signatures allowed us to identify the most commonly modulated pathways by both drugs. In brief, we observed predominantly upmodulation of transcripts related to PI3K-Akt, NF-kappaB, and tumor necrosis factor (TNF) signaling pathways as well as transcripts related to negative regulation of apoptotic process among others. In addition, hematopoietic colony-forming cell assays corroborate the G-CSF phenotypic effects over UBC-derived MNCs. In conclusion, our study suggests that G-CSF impacts UBC-derived cells through the modulation of several signaling pathways associated with cell survival, migration, and proliferation. The concordance observed between biosimilar and innovator G-CSF emphasizes their similarity in regards to their specificity and biological responses.


Blood ◽  
2009 ◽  
Vol 114 (22) ◽  
pp. 2159-2159
Author(s):  
Bryan P Spencer ◽  
Indreshpal Kaur ◽  
Patrick Fong ◽  
Safa Karandish ◽  
Nirmali Ponweera ◽  
...  

Abstract Abstract 2159 Poster Board II-136 INTRODUCTION: Many procedures for manufacturing clinical products for cellular therapy involve enrichment of mononuclear cells (MNC). The most common procedure is density gradient separation. Disadvantages of this procedure are low yield of cells especially with cryopreserved products and the open events during processing. We evaluated use of the Purecell” Select System (PALL Medical, NY) for enriching MNC from cryopreserved Cord Blood Units (CBU). METHODS: Initial experiments were performed to optimize the system for recoveries of total nucleated cells (TNC), MNC, neutrophils, lymphocytes, monocytes and CD34+/CD3+ cells. We evaluated thaw/predilute/filter vs thaw/filter, starting volumes (30 — 95mls) and three different methods for harvesting cells from the filter (standard method, input bag rinse and harvest port rinse). Once conditions were optimized, cryopreserved CBU were thawed and split into two fractions. One half of the product was diluted and processed on the Purecell” Select System. The other half was washed and ficolled. The MNC fraction was CD3+ enriched using CD3/28 beads and then cultured with rIL-2 (200 units/ml) for 14 days. The absolute number of CD3+ cells post culture, fold expansion and viabilities of these cells were determined. RESULTS: The Purecell” procedure was 15 times faster than the ficoll method. Optimal volume to load onto the filter was 50ml. When MNC were harvested by the three recommended procedures, there was no difference in recoveries of TNC, MNC, CD34+ and CD3+ cells and neutrophils. However, the lymphocyte and monocyte recoveries were higher (p<0.05 and p=0.001) when harvested with Input bag rinse compared to the standard procedure. Monocyte recoveries were also higher with the harvest port rinse (p=0.004) when compared to the standard procedure. The direct comparison studies of the two MNC enrichment systems demonstrated that the Purecell” Select System gave significantly higher recovery of TNC (p= 0.003), MNC (p=0.029), CD34+ cells (p<0.001) and granulocytes (p<0.001). There was no statistical difference in T cell recoveries, however, there was a significant difference in the recovery of T cells after CD3/28 enrichment.. Interestingly, T cells began to proliferate earlier from the PALL system compared to ficoll isolated T cells (day 4 vs day 7). Although the fold expansion was greater for the ficolled prepared cells, the absolute numbers of T cells obtained after 14 days of culture with rIL-2 was greater for the Purecell Select System in all experiments. The viabilities of the cells from both cultures were comparable. CONCLUSIONS: PALL's Purecell” Select System can be used for clinical processing since it is a functionally closed system. The advantage of this system compared to the ficoll method are the reduced time for processing, increase yield in T cells (post processing), the earlier expansion time These benefits result in an increase in absolute number of T cells in post culture. A clinical trial using this system is about to be initiated. Disclosures: Karandish: Pall Medical: Employment. McMannis:Pall Medical: Research Funding.


Blood ◽  
1995 ◽  
Vol 86 (10) ◽  
pp. 3745-3753 ◽  
Author(s):  
QL Hao ◽  
AJ Shah ◽  
FT Thiemann ◽  
EM Smogorzewska ◽  
GM Crooks

We present cell cycling and functional evidence that the CD34+CD38- immunophenotype can be used to define a rare and primitive subpopulation of progenitor cells in umbilical cord blood. CD34+CD38- cells comprise 0.05% +/- 0.08% of the mononuclear cells present in cord blood. Cell cycle analysis with the fluorescent DNA stain 7- aminoactinomycin D showed that the percentage of CD34+ cells in cycle directly correlated with increasing CD38 expression. CD34+CD38- cord blood cells were enriched for long-term culture-initiating cells (LTCIC; cells able to generate colony-forming unit-cells [CFU-C] after 35 to 60 days of coculture with bone marrow stroma) relative to CD34+CD38- cells. In an extended LTCIC assay, CD34+CD38- cells were able to generate CFU-C between days 60 and 100, clearly distinguishing them from CD34+CD38+ cells that did not generate CFU-C beyond day 40. When plated as single cells, onset of clonal proliferation was markedly delayed in a subpopulation of CD34+CD38- cells; clones (defined as = 100 cells) appeared after 60 days of culture in 2.9% of CD34+CD38- cells. In contrast, 100% of CD34+CD38+ cells formed clones by day 21. Although the CD34+CD38- immunophenotype defines highly primitive populations in both bone marrow and cord blood, important functional differences exist between the two sources. CD34+CD38- cord blood cells have a higher cloning efficiency, proliferate more rapidly in response to cytokine stimulation, and generate approximately sevenfold more progeny than do their counterparts in bone marrow.


Blood ◽  
2004 ◽  
Vol 104 (11) ◽  
pp. 4206-4206
Author(s):  
Angela C. Luzo ◽  
Adriana S. Duarte ◽  
Anderson F. Cunha ◽  
Dulcineia M. Albuquerque ◽  
Barbara C. Monte-Mor ◽  
...  

Abstract Hematopoietic stem/progenitor cells with their dual ability for self-renewal, expansion and multilineage differentiation constitute an essential component of hematopoietic transplantation. These qualities, together with their transfection capacity make them useful for gene therapy also. Umbilical cord blood and bone marrow represent efficient sources of these cells. Cord blood stem cells are more primitive than bone marrow cells. They have a greater capacity to self-renewal, proliferation, expansion, multilineage differentiation and are better transfected. They can also be transplanted without complete HLA compatibility. Some recent studies are demonstrating that they could have a better trafficking and homing than the others, which could promote an efficient development and proliferation in the marrow microenvironment and better recovering when transplanted. However, these mechanisms of migration/cell adhesion and cell cycle are not fully understood. The aim of this study is to compare cord blood mononuclear cells with the existing bone marrow mononuclear cells library (esage database) by means of serial analysis of gene expression in order to investigate if there are any differences that could explain those findings. We obtained, by sequencing the SAGE library of cord blood cells, a total of 44.924 tags representing 15.519 unique tags. These unique tags were distribute as: known genes= 7772; sequence predicted or annotated=3935; no matches= 3812. The bone marrow library showed total tags=36577 being unique tags=13075: known genes= 6711; sequence predicted or annotated = 3371; no matches= 2993. These genes were annotated using Gene Ontology Consortium and the distribution of the categories in both libraries was similar. The different expression genes, selected using the fold difference (fold≥5 and ≤ 5), were 238. In order to validate the library, we tested 12 different expressed genes by real time PCR. The first six of them (SMARCC2, CDC25B, S100-8, alpha-globin, beta-globin, gamma-globin) confirmed the SAGE results. The more expressed cord blood genes were SMARCC2, CDC25B and gamma-globin. Beta-globin, and S100-8 were more expressed in bone marrow. The alpha- globin result demonstrated almost the same result between cord blood and bone marrow. CDC25B is a member of the CDC25, family of phosphatases, it activates cyclin dependent kinase CDC2 which is required for entry into mitoses. SMARCC2 is a member of SWI/SNF family proteins which display helicase and ATPase activities. It is responsible for regulation of transcription of certain genes by altering the chromatin structure around those genes. S100-8 is a calcium binding protein with a prominent role in leukocyte trafficking. It stimulates shedding of L-selectin, which is also involved in cell adhesion. It makes a role in inflammatory process towards neutrophil migration to inflammatory sites. The alpha, beta and gamma globin results were those expected based on the ontogeny. The differential gene expression here obtained could be an important tool to guide new functional studies regarding the molecular mechanisms of homing, cell cycle and cell adhesion of cord blood cells.


Blood ◽  
1996 ◽  
Vol 87 (8) ◽  
pp. 3344-3350 ◽  
Author(s):  
S Saito ◽  
T Morii ◽  
H Umekage ◽  
K Makita ◽  
K Nishikawa ◽  
...  

Lymphocytes in umbilical cord blood and neonatal peripheral blood have been shown to have less ability in an immune reaction. In our present experimental approach to address this issue, we made use of the cord blood of full-term birth infants to investigate the expression of the interleukin- 2 receptor gamma (IL-2Rgamma) chain that is shared with receptors for IL-4, IL-7, IL-9, and IL-15 as well as IL-2. The gamma chain expression in cord blood lymphocytes was about one-third that in the lymphocytes of adults, whereas no significant difference between cord blood and adult monocytes was observed. A reduced expression of the gamma chain was observed in all of the CD4+ T cells, CD8+ T cells, gamma-delta T cells, B cells, CD16+ natural killer (NK) cells, and CD56(bright) NK cells of the cord blood lymphocytes. The reduced gamma chain expression reached two-thirds of that in adults after 3 days of culture in vitro and in infants 3 days after birth, thus implying that the increase in the gamma chain may significantly contribute to the prevention of neonatal infection.


Blood ◽  
2006 ◽  
Vol 108 (11) ◽  
pp. 5212-5212 ◽  
Author(s):  
Maria Felicitas N. Rodriguez ◽  
Alexandra C. Senegaglia ◽  
Carmem K. Rebelatto ◽  
Daniela V. Pilonetto ◽  
Samira Fadel ◽  
...  

Abstract Cord blood transplantation (CBT) has been considered a treatment modality to hematologic malignancies, marrow failure syndromes and genetic diseases. It has advantages such as immediate availability, lower risk of transmitting infections and higher tolerance to HLA disparities compared to bone marrow. The major drawbacks are slow hematopoietic recovery and a high incidence of graft failure or delayed engraftment, as a result of low stem progenitors (CD34+cells) yields. This study aimed at investigating if there was correlation between engraftment and number of total nucleated cells (TNC), number of CD34+ cells as well as number of granulocyte/monocyte colony forming units (CFU-GM) and erythroid burst forming units (BFU-E). Eighty four patients (age 0 to 55, M=7) with a variety of diseases (10 AML, 12 ALL, 31 Fanconi Anemia, 1 Hodgkin Lymphoma, 3 JMML, 3 Blackfan Diamond, 6 Wiskott Aldrich, 1 Krabbe Disease, 2 SAA, 4 CML, 3 Osteopetrosis, 3 SCID, 1 Diskeratosis Congenita, 1 Adrenoleukodystrophy, 1 Thalassemia Major, 1 Thrombocytopenia w/chromosome 7 monossomy) who underwent CBT were included in this study; 47.6% (40/84) of these patients had graft failure while 52.4% (44/84) showed successful engraftment. Regarding HLA-A, B and DRB1 matching, 20.2% (17/84) recipient/donor pairs were 6/6, 42.9% (36/84) were 5/6, 35.7% (30/84) were 4/6 and 1.2% (1/84) was 3/4. TNC and CD34+ cell counts in addition to hematopoeitic progenitor cells (CFU-GM and BFU-E) testing were performed in cord blood products after thawing and washing procedures. CD34+cells were analyzed by two color technique using FACS Calibur (San Jose, CA) and absolute counts were assessed by two-platform ISHAGE method. Colony assays for BFU-E and CFU-GM were performed by plating 3×104 mononuclear cells in methylcellulose medium with human recombinant cytokines (Methocult GF, Stem Cell Technologies); cultures were kept at 37°C in humidified incubator with 5% CO2 and after 14 days colonies were scored under an inverted microscope. Comparison of pre-thawing TNC [M=1.34×109(0.56–5.00)] and CD34+ [M=2.61×105/Kg (0.39–25.48)] cell counts, provided by cord blood banks, with post-thawing TNC [M=1.09×109(0.36–4.20)] and CD34+ [M=1.3×105/Kg(0.06–9.8)], performed by Wilcoxon signed rank test, showed a significant decrease in both cases (P<0.001). Descriptive statistics were tabulated separately for the number of TNC, number of CD34+ cells, numbers of CFU-GM and BFU-E colonies by whether patient engrafted or had graft failure, and comparison of the median cell counts between these two groups of patients was done by Wilcoxon Mann-Whitney rank sum test. TNC and CD34+ cell counts did not show significant difference between patients with graft failure [M=1.08×109(0.36–4.2) and M=1.28×105/Kg(0.09–9.8), respectively] and those who engrafted [M=1.12×109(0.46–2.49) and M=1.41×105/Kg(0.06–6.0), respectively]. Yet, the CFU-GM counts were significantly higher (P=0.005) for patients who engrafted [M=0.079×105(0.0017–0.75)] than for those who did not [M=0.04×105(0.0009–0.50)]. Although BFU-E counts did not differ with statistical significance between engrafted versus graft failure patients (P=0.059), higher counts could be observed in the group of patients who had successful engraftment. These data suggest that results of counts of CFU-GM colonies have a predictive value for engraftment in our center.


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