scholarly journals Phenotype and Function of CD25-Expressing B Lymphocytes Isolated from Human Umbilical Cord Blood

2011 ◽  
Vol 2011 ◽  
pp. 1-9 ◽  
Author(s):  
Sylvie Amu ◽  
Mikael Brisslert
Keyword(s):  
B Cells ◽  
Cord Blood ◽  
B Cell ◽  
Cell Subset ◽  
Cell Populations ◽  
Human Umbilical Cord Blood ◽  
Human Umbilical Cord ◽  
Common Denominator ◽  
Human Cord Blood ◽  
And Function

Background. We have shown that approximately 30% of human peripheral blood B-cells express CD25. B cells expressing CD25 display a mature phenotype belonging to the memory B-cell population and have a better proliferative and antigen-presenting capacity. The aim of the present study was to characterize the CD25-expressing subset of B cells in human cord blood.Material and Methods. Mononuclear cell fraction from human cord blood (n=34) and peripheral adult blood (n=22) was sorted into CD20+CD25+and CD20+CD25-B-cell populations. Phenotype and function of these B-cell populations were compared using flow cytometry, proliferation, cytokine production, and immunoglobulin secretion.Results. CD25-expressing B cells are a limited population of cord blood mononuclear cells representing 5% of the CD20+B cells. They are characterised by high expression of CD5 in cord blood and CD27 in adult blood. CD25-expressing B cells express a functional IL-2 receptor and high levels of CC-chemokine receptors and spontaneously produce antibodies of IgG and IgM subclass.Conclusions. CD25 expression is a common denominator of a specific immunomodulatory B-cell subset ready to proliferate upon IL-2 stimulation, possibly ready to migrate and home into the peripheral tissue for further differentiation/action.

scholarly journals Human Umbilical Cord Blood-Derived Serum for Culturing the Supportive Feeder Cells of Human Pluripotent Stem Cell Lines

2016 ◽  
Vol 2016 ◽  
pp. 1-12 ◽  
Author(s):  
Ruttachuk Rungsiwiwut ◽  
Praewphan Ingrungruanglert ◽  
Pranee Numchaisrika ◽  
Pramuan Virutamasen ◽  
Tatsanee Phermthai ◽  
...  
Keyword(s):  
Stem Cells ◽  
Cord Blood ◽  
Culture System ◽  
Extended Period ◽  
Population Doubling ◽  
Human Umbilical Cord Blood ◽  
Human Umbilical Cord ◽  
Feeder Cells ◽  
Human Cord Blood ◽  
Stem Cell Lines

Although human pluripotent stem cells (hPSCs) can proliferate robustly on the feeder-free culture system, genetic instability of hPSCs has been reported in such environment. Alternatively, feeder cells enable hPSCs to maintain their pluripotency. The feeder cells are usually grown in a culture medium containing fetal bovine serum (FBS) prior to coculture with hPSCs. The use of FBS might limit the clinical application of hPSCs. Recently, human cord blood-derived serum (hUCS) showed a positive effect on culture of mesenchymal stem cells. It is interesting to test whether hUCS can be used for culture of feeder cells of hPSCs. This study was aimed to replace FBS with hUCS for culturing the human foreskin fibroblasts (HFFs) prior to feeder cell preparation. The results showed that HFFs cultured in hUCS-containing medium (HFF-hUCS) displayed fibroblastic features, high proliferation rates, short population doubling times, and normal karyotypes after prolonged culture. Inactivated HFF-hUCS expressed important genes, including Activin A, FGF2, and TGFβ1, which have been implicated in the maintenance of hPSC pluripotency. Moreover, hPSC lines maintained pluripotency, differentiation capacities, and karyotypic stability after being cocultured for extended period with inactivated HFF-hUCS. Therefore, the results demonstrated the benefit of hUCS for hPSCs culture system.

Human cord blood-derived cells attain neuronal and glial features in vitro

2002 ◽  
Vol 115 (10) ◽  
pp. 2131-2138 ◽  
Author(s):  
L. Bużańska ◽  
E. K. Machaj ◽  
B. Zabłocka ◽  
Z. Pojda ◽  
K. Domańska-Janik
Keyword(s):  
Stem Cells ◽  
Neural Stem Cells ◽  
Cord Blood ◽  
Human Umbilical Cord Blood ◽  
Specific Cell ◽  
Human Umbilical Cord ◽  
Neurofilament Protein ◽  
Adhesive Properties ◽  
Human Cord Blood

Neural stem cells are clonogenic, self-renewing cells with the potential to differentiate into brain-specific cell lines. Our study demonstrates that a neural-stem-cell-like subpopulation can be selected and expanded in vitro by the use of human umbilical cord blood cells, which are a relatively easily available starting material. Through a combination of antigen-driven magnetic cell sorting and subfractionation according to cell surface adhesive properties, we have isolated a clonogenic fraction devoid of hematopoietic or angiogenetic properties but with relatively high self-renewal potency. The resulting clones express nestin, a neurofilament protein that is one of the most specific markers of multipotent neural stem cells. In the presence of selected growth factors or in the rat brain co-culture system, the progeny of these cells can be oriented towards the three main neural phenotypes: neurons,astroglia and oligodendroglia. The cells show high commitment (about 30% and 40% of the population) to neuronal and astrocytic fate, respectively. Interestingly, upon differentiation, the neural-type precursor cells of cord blood origin also give rise to a relatively high proportion of oligodendrocytes — 11% of the total population of differentiating cells.

scholarly journals B-cell differentiation following autologous, conventional, or T-cell depleted bone marrow transplantation: a recapitulation of normal B-cell ontogeny

Blood ◽  
1990 ◽  
Vol 76 (8) ◽  
pp. 1647-1656 ◽  
Author(s):  
TN Small ◽  
CA Keever ◽  
S Weiner-Fedus ◽  
G Heller ◽  
RJ O'Reilly ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Cell Differentiation ◽  
Bone Marrow Transplantation ◽  
B Cells ◽  
Cord Blood ◽  
B Cell ◽  
First Year ◽  
B Cell Differentiation ◽  
And Function ◽  
Cell Ontogeny

Abstract The circulating lymphocytes of 88 consecutive patients following autologous, conventional, or T-cell depleted bone marrow transplantation were serially analyzed for B-cell surface antigen expression and function. In the majority of patients, except for those who developed chronic graft-versus-host disease, the number of circulating CD20+ B cell normalized by the fourth posttransplant month. The earliest detectable B cells normally expressed HLA-DR, CD19, surface immunoglobulin (slg), CD21, Leu-8, and lacked expression of CD10 (CALLA). In addition, the circulating B cells expressed CD1c, CD38, CD5, and CD23 for the first year following transplant, antigens that are normally expressed on a small percentage of circulating B cells in normal adults, but highly expressed on cord blood B cells. Similar to cord blood B cells, patient B cells isolated during the first year following transplant, proliferated normally to Staphylococcus aureus Cowan strain I (SAC), and produced IgM, but minimal or no IgG when stimulated with pokeweed mitogen and SAC, unlike normal adult B cells that produce both. The similar phenotype and function of posttransplant and cord blood B cells, and their similar rate of decline in patients and normal children adds further evidence to support the hypothesis that B-cell differentiation posttransplant is recapitulating normal B-cell ontogeny.

scholarly journals Functional properties of cells obtained from human cord blood CD34+ stem cells and mouse cardiac myocytes in coculture

2008 ◽  
Vol 294 (4) ◽  
pp. H1541-H1549 ◽  
Author(s):  
Alessia Orlandi ◽  
Francesca Pagani ◽  
Daniele Avitabile ◽  
Giuseppina Bonanno ◽  
Giovanni Scambia ◽  
...  
Keyword(s):  
Stem Cells ◽  
Cord Blood ◽  
Functional Properties ◽  
Fluorescent Protein ◽  
Cell Populations ◽  
Human Umbilical Cord Blood ◽  
Human Umbilical Cord ◽  
Rt Pcr ◽  
Hematopoietic Stem ◽  
Human Specific

Prior in vitro studies suggested that different types of hematopoietic stem cells may differentiate into cardiomyocytes. The present work examined whether human CD34+ cells from the human umbilical cord blood (hUCB), cocultured with neonatal mouse cardiomyocytes, acquire the functional properties of myocardial cells and express human cardiac genes. hUCB CD34+ cells were cocultured onto cardiomyocytes following an infection with a lentivirus-encoding enhanced green fluorescent protein (EGFP). After 7 days, mononucleated EGFP+ cells were tested for their electrophysiological features by patch clamp and for cytosolic [Ca2+] ([Ca2+]i) homeostasis by [Ca2+]i imaging of X-rhod1-loaded cells. Human Nkx2.5 and GATA-4 expression was examined in cocultured cell populations by real-time RT-PCR. EGFP+ cells were connected to surrounding cells by gap junctions, acquired electrophysiological properties similar to those of cardiomyocytes, and showed action potential-associated [Ca2+]i transients. These cells also exhibited spontaneous sarcoplasmic reticulum [Ca2+]i oscillations and the associated membrane potential depolarization. However, RT-PCR of both cell populations showed no upregulation of human-specific cardiac genes. In conclusion, under our experimental conditions, hUCB CD34+ cells cocultured with murine cardiomyocytes formed cells that exhibited excitation-contraction coupling features similar to those of cardiomyocytes. However, the expression of human-specific cardiac genes was undetectable by RT-PCR.

scholarly journals Characterization of glycoprotein IIb/IIIa-positive cells in human umbilical cord blood: their potential usefulness as megakaryocyte progenitors

Blood ◽  
1992 ◽  
Vol 79 (2) ◽  
pp. 347-355
Author(s):  
D Zucker-Franklin ◽  
JS Yang ◽  
G Grusky
Keyword(s):  
Stem Cells ◽  
Cord Blood ◽  
Experimental Studies ◽  
Cell Types ◽  
Mononuclear Leukocytes ◽  
Human Umbilical Cord Blood ◽  
Human Umbilical Cord ◽  
Phenotypic Analysis ◽  
Human Cord Blood ◽  
Hematopoietic Stem

A need for hematopoietic stem cells, particularly cells destined to enter the megakaryocyte (MK) series, prompted phenotypic analysis of mononuclear leukocytes in human cord blood. To this end, immunohistochemical, flow cytometric, and ultrastructural techniques were used. The immunogold silver enhancement method (IGS) for the detection of the MK-specific glycoprotein (GP) IIb/IIIa epitopes combined with a monocyte-specific stain for alpha-naphthyl butyrate esterase proved to be superior to flow cytometry (FACS) for precise quantitation of cell types in each sample. Immunoelectron microscopy afforded a description of distinctions between precursors bearing GPIIb/IIIa epitopes and other stem cells of the myeloid series. The number of presumed MK progenitors was surprisingly high, averaging 1.8% +/- 1.3% (range, 0.2% to 4.6%) by IGS and 4.1% +/- 3.0% (range, 0.2% to 9.3%) by FACS analysis. The occurrence of GPIIb/IIIa-positive denuded MK nuclei in cord blood was of interest, but was too small to affect these data. These observations should advocate a greater use of cord blood for restitution of MK/platelet-lineage-depleted patients as well as for experimental studies concerned with MK differentiation.

scholarly journals In Vitro Growth of Granulocytic Colonies From Circulating Cells in Human Cord Blood

Blood ◽  
1974 ◽  
Vol 43 (3) ◽  
pp. 357-361 ◽  
Author(s):  
Søren Knudtzon
Keyword(s):  
Cord Blood ◽  
Peripheral Blood ◽  
Blood Cells ◽  
Newborn Infants ◽  
Peripheral Blood Cell ◽  
Human Umbilical Cord Blood ◽  
Human Umbilical Cord ◽  
Human Cord Blood ◽  
Cord Blood Cells

Abstract Human umbilical cord blood cells from 26 newborn infants and peripheral blood cells from 18 adults were cultured in vitro by using the agar-gel method of human hemopoietic cell culture. An increased concentration of colony-forming cells was seen in the cord blood cultures. Between 17 and 385 colonies, with a mean of 122, were formed in these cultures per 2 x 105 nucleated cells plated. The peripheral blood cell cultures from adults gave rise to 0-11 colonies, with a mean of 3, per 2 x 105 nucleated cells plated. The average number of cells per colony was 1000-1500 cells after 14 days of culture, predominantly granulocytic.

Augmented Effects on Neovascularization by Mixing the Conditioned Medium from Early EPCs into Late EPCs Derived from Human Cord Blood.

Blood ◽  
2006 ◽  
Vol 108 (11) ◽  
pp. 3936-3936
Author(s):  
Eun-Sun Yoo ◽  
Jee-Young Ahn ◽  
Yun-Kyung Bae ◽  
Jee Hyun Kong ◽  
Sang Min Lee ◽  
...  
Keyword(s):  
Cord Blood ◽  
Conditioned Medium ◽  
Mononuclear Cells ◽  
Ex Vivo ◽  
Tube Formation ◽  
Marker Genes ◽  
Human Umbilical Cord Blood ◽  
Human Umbilical Cord ◽  
Migration Ability ◽  
Human Cord Blood

Abstract Recently, we have reported on two different types of EPCs from human umbilical cord blood(ASH 2005, Abstract #1706). These EPCs had different biologic properties in angiogenic capabilities during the HCB ex vivo expansion. In this present study, the aim is to examine the synergism by mixing the conditioned medium from the early EPCs into the late EPCs on neovascularization. The mononuclear cells from the HCB were cultures using an EGM-2 medium with VEGF, IGF-1 and FGF for 21 days. We found early spindle-shaped cells(early EPCs), which were grown in the first week of the culture and late cobblestone shaped cells(late EPCs) which peaked in their growth in the third week of the culture. First, we compared the two types of cells in terms of phenotypic expressions and migration ability. Next, we examined their proliferation capacity and tube formations in the Matrigel plate under the conditions that the early outgrowing cells-contained medium was added to the cobblestone shaped cells. The late-appearing cobblestone shaped cells were positive for VEGFR2, VE-cadherin, CD31, CD34 and CXCR-4 but not for CD14 and CD54. These late outgrowing cells expressed high levels of mRNA on the endothelial marker genes and effectively formed capillary tubes in the Matrigel plates. The early spindle cells excreted more angiogenic cytokines and had more migratory ability. When the early spindle-shaped cell-conditioned medium was added to the late cobblestone shaped outgrowing cells, significantly higher proliferation and tube formation measured by the area and length of tubes were found. These results suggested that the two types of cells raised with different biologic properties during the ex vivo HCB expansion and the angiogenic capacity of late EPCs were augmented by a mutual interaction via the excreted cytokines from early EPCs. These finding may have potential applications for a “cell therapy” in situations such as vascular injuries (ie, hindlimb ischemia/myocardial infarction). Murine models are being tested to see whether the injections of two different EPCs will result in synergic noevascularization in our Lab.

Mir-125a Confers Multi-Lineage Long-Term Repopulating Stem Cell Activity to Human Hematopoietic Committed Progenitors

Blood ◽  
2015 ◽  
Vol 126 (23) ◽  
pp. 900-900 ◽  
Author(s):  
Eric R. Lechman ◽  
Karin G. Hermans ◽  
Erwin M. Schoof ◽  
Aaron Trotman-Grant ◽  
Stephanie M Dobson ◽  
...  
Keyword(s):  
Stem Cell ◽  
B Cell ◽  
Conflicts Of Interest ◽  
Cell Activity ◽  
Lymphoid Cells ◽  
Cell Populations ◽  
Human Umbilical Cord Blood ◽  
Human Umbilical Cord ◽  
Self Renewal ◽  
Hematopoietic Stem

Abstract Recent studies have shown that several miRNA are differentially expressed in hematopoietic stem cells (HSC) and involved in regulating self-renewal, pointing to a new axis of epigenetic control of HSC function. Murine studies have documented a role for miR-125a in regulating HSC as miR-125a enforced expression augments self-renewal. We examined whether these attributes are evolutionarily conserved within human hematopoiesis. Lentiviral vectors over-expressing miR-125a (miR-125OE) were developed and HSC function was investigated using xenotransplantation of CD34+ CD38- human umbilical cord blood (CB) hematopoietic stem and progenitor cells (HSPCs). miR-125OE resulted in significantly increased human bone marrow (BM) chimerism at 12 and 24 weeks post-transplantation and splenomegaly. Within enlarged spleens, there were significantly increased proportions of CD34+CD19+CD10+CD20-B lymphoid cells suggesting a partial B cell differentiation block at the pro-B cell stage. In the BM, CD41+ megakaryocytes, GlyA+ erythroid and CD3+ T cell populations were significantly expanded. Within the primitive compartment, multi-lymphoid progenitors (MLP) were massively expanded by 12 weeks, followed by a combined reduction of immuno-phenotypic HSC and multi-potent progenitors (MPP) by 24 weeks. Given this loss of immuno-phenotypic HSC, we wondered whether stem cell function was compromised in vivo. Secondary transplantation with limiting dilution (LDA) revealed that stem cell frequencies were increased by 4.5 fold in miR-125OE recipients. Using lentivirus sponge-mediated inhibition of miR-125 (miR-125KD) in CD34+CD38-human CB, we were able to directly link these effects to miR-125: B cells increased at the expense of T cells; immuno-phenotypic HSC increased with a concomitant loss of MLP; and functional HSC were decreased by 2.5 fold using secondary LDA assays. Together, these data strongly suggest that miR-125a expression levels regulate human HSC self-renewal and lineage commitment. Since HSC frequency increased so substantially upon miR-125OE, we asked whether more committed cell populations might also be endowed with enhanced self-renewal. Highly purified populations of HSC, MPP and MLP and CD34+CD38+ committed progenitors were transduced and transplanted cells into xenografts. Unexpectedly, miR-125OE transduced CD34+CD38+ progenitors produced a substantial graft after 12 weeks. Control transduced CD34+CD38+ cells did not engraft and only control transduced HSC generated a disseminating graft in recipient mice. miR-125OE transduced HSC and MPP generated robust engraftment, while MLP did not. In all cases, xenografts generated by CD34+CD38+ and MPP transduced with miR-125OE showed multi-lineage repopulation. Moreover, the miR-125OE grafts from CD34+CD38+ and MPP recipients were durable as secondary transplantation generated multi-lineage grafts for at least 20 weeks in 5/7 and 6/10 recipients, respectively; no control transduced groups generated secondary grafts. Thus, the enhancement of self-renewal by enforced expression of miR-125a occurs not only in HSC, but also in MPP and to an as yet unidentified subpopulation within the CD34+38+ committed progenitor compartment. Using protein mass spectrometry, we identified and validated a miR-125a target network in CD34+ CB that normally functions to restrain self-renewal in more committed progenitors. Together, our data suggest that increased miR-125a expression can endow an HSC-like program upon a selected set of non-self-renewing hematopoietic progenitors. Our findings offer the innovative potential to use MPP with enhanced self-renewal to augment limited sources of HSC to improve clinical outcomes. Disclosures No relevant conflicts of interest to declare.

scholarly journals B-cell differentiation following autologous, conventional, or T-cell depleted bone marrow transplantation: a recapitulation of normal B-cell ontogeny

Blood ◽  
1990 ◽  
Vol 76 (8) ◽  
pp. 1647-1656 ◽  
Author(s):  
TN Small ◽  
CA Keever ◽  
S Weiner-Fedus ◽  
G Heller ◽  
RJ O'Reilly ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Cell Differentiation ◽  
Bone Marrow Transplantation ◽  
B Cells ◽  
Cord Blood ◽  
B Cell ◽  
First Year ◽  
B Cell Differentiation ◽  
And Function ◽  
Cell Ontogeny

The circulating lymphocytes of 88 consecutive patients following autologous, conventional, or T-cell depleted bone marrow transplantation were serially analyzed for B-cell surface antigen expression and function. In the majority of patients, except for those who developed chronic graft-versus-host disease, the number of circulating CD20+ B cell normalized by the fourth posttransplant month. The earliest detectable B cells normally expressed HLA-DR, CD19, surface immunoglobulin (slg), CD21, Leu-8, and lacked expression of CD10 (CALLA). In addition, the circulating B cells expressed CD1c, CD38, CD5, and CD23 for the first year following transplant, antigens that are normally expressed on a small percentage of circulating B cells in normal adults, but highly expressed on cord blood B cells. Similar to cord blood B cells, patient B cells isolated during the first year following transplant, proliferated normally to Staphylococcus aureus Cowan strain I (SAC), and produced IgM, but minimal or no IgG when stimulated with pokeweed mitogen and SAC, unlike normal adult B cells that produce both. The similar phenotype and function of posttransplant and cord blood B cells, and their similar rate of decline in patients and normal children adds further evidence to support the hypothesis that B-cell differentiation posttransplant is recapitulating normal B-cell ontogeny.

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