Phenotypic Characterization of Immunomagnetically Purified Umbilical Cord Blood CD34+ Cells

Belvedere, Ornella; Feruglio, Cristina; Malangone, W.; Bonora, Maria Letizia; Donini, Annibale; Dorotea, Laura; Tonutti, Elio; Rinaldi, Cristina; Pittino, Marco; Baccarani, Michele; Frate, G. Del; Biffoni, Franco; Sala, Pierguido; Hilbert, David M.; Degrassi, A.

doi:10.1006/bcmd.1999.0239

Cited by 22 publications

(23 citation statements)

References 18 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…Fast hemopoietic recovery after transplantation of adult mobilized peripheral blood (mPB) HSC, which consists mainly of CD38 ϩ cells (8), suggests a role for CD34 ϩ 38 ϩ cells, especially during the early posttransplant phase. These cells may be important in shortening the aplastic phase.…”

Section: T He True Hemopoietic Stem Cell (Hsc)mentioning

confidence: 99%

“…In vitro data have shown that the CD34 ϩ CD38 ϩ fraction contains colony-forming cells and longterm culture-initiating cells and that these develop more quickly but are more quickly exhausted than from CD34 ϩ 38 Ϫ cells (5, 9, 10). Transplantation of mPB HSC, which mainly consist of CD34 ϩ CD38 ϩ cells (8), results in fast reconstitution, and it is reasonable to assume that these early developing cells are the progeny of the bulk of the cells. Moreover, in vitro expansion with both early-and late-acting growth factors of purified HSC resulted in significant shortening of the aplastic phase.…”

mentioning

confidence: 99%

See 1 more Smart Citation

Both CD34+38+ and CD34+38− Cells Home Specifically to the Bone Marrow of NOD/LtSZ scid/scid Mice but Show Different Kinetics in Expansion

Kerre

Smet

Smedt

et al. 2001

The Journal of Immunology

View full text Add to dashboard Cite

Human hemopoietic stem cells (HSC) have been shown to engraft, differentiate, and proliferate in the hemopoietic tissues of sublethally irradiated NOD/LtSZ scid/scid (NOD/SCID) mice. We used this model to study homing, survival, and expansion of human HSC populations from different sources or phenotype. We observed that CD34+ cells homed specifically to bone marrow (BM) and spleen, but by 3 days after injection, survived only in the BM. These BM-homed CD34+ cells proliferated intensively and gave rise to a 12-fold, 5.5-fold, and 4-fold expansion in 3 days for umbilical cord blood, adult mobilized peripheral blood, and adult BM-derived cells, respectively. By injection of purified subpopulations, it was demonstrated that both CD34+38+ and CD34+38− umbilical cord blood HSC homed to the BM and expanded. Importantly, kinetics of expansion were different: CD34+38+ cells started to increase in cell number from day 3 onwards, and by 4 wk after injection, virtually all CD34+ cells had disappeared. In contrast, CD34+38− cells remained quiescent during the first week and started to expand intensively from the third week on. In this paper, we have shown that homing, survival, and expansion of stem cells are three independent phenomena important in the early phase of BM engraftment and that kinetics of engraftment differ between CD34+38+ and CD34+38− cells.

show abstract

Section: T He True Hemopoietic Stem Cell (Hsc)mentioning

confidence: 99%

mentioning

confidence: 99%

Both CD34+38+ and CD34+38− Cells Home Specifically to the Bone Marrow of NOD/LtSZ scid/scid Mice but Show Different Kinetics in Expansion

Kerre

Smet

Smedt

et al. 2001

The Journal of Immunology

View full text Add to dashboard Cite

show abstract

“…However, a better knowledge and standardization of the phenotype of umbilical cord blood CD34 + cells is critical since HUCB volume is limited (13). The present study aimed to contribute to the characterization of CD34 + cells from umbilical cord blood, analyzing their phenotype and behavior before and after culture with different combinations of growth factors.…”

Section: Discussionmentioning

confidence: 99%

“…This might explain the successful clinical use of HUCB even when a small number of cells is used, making the presence of these antigens candidate predictive parameters for clinical use of HUCB samples (13).…”

Section: Discussionmentioning

confidence: 99%

“…There is evidence that cord blood, bone marrow and peripheral blood-derived HSCs are highly heterogeneous for a number of antigens useful for HSC enumeration by flow cytometry (6). HLA-DR is expressed in the majority of HUCB (4,13) and peripheral blood (13,19) CD34 + cells. While De Bruyn et al (21) showed that the co-expression of CD34 with HLA-DR was not significantly different in HUCB and bone marrow (86.3 ± 2.7 and 92.7 ± 5.1%, respectively), Cho et al (22) showed that CD34 + HLA-DR + cell frequencies did not differ significantly between those two compartments and MNCs.…”

Section: Discussionmentioning

confidence: 99%

See 1 more Smart Citation

Immunophenotype of hematopoietic stem cells from placental/umbilical cord blood after culture

Pranke

Hendrikx

Debnath

et al. 2005

Braz J Med Biol Res

View full text Add to dashboard Cite

Identification and enumeration of human hematopoietic stem cells remain problematic, since in vitro and in vivo stem cell assays have different outcomes. We determined if the altered expression of adhesion molecules during stem cell expansion could be a reason for the discrepancy. CD34+CD38- and CD34+CD38+ cells from umbilical cord blood were analyzed before and after culture with thrombopoietin (TPO), FLT-3 ligand (FL) and kit ligand (KL; or stem cell factor) in different combinations: TPO + FL + KL, TPO + FL and TPO, at concentrations of 50 ng/mL each. Cells were immunophenotyped by four-color fluorescence using antibodies against CD11c, CD31, CD49e, CD61, CD62L, CD117, and HLA-DR. Low-density cord blood contained 1.4 ± 0.9% CD34+ cells, 2.6 ± 2.1% of which were CD38-negative. CD34+ cells were isolated using immuno-magnetic beads and cultured for up to 7 days. The TPO + FL + KL combination presented the best condition for maintenance of stem cells. The total cell number increased 4.3 ± 1.8-fold, but the number of viable CD34+ cells decreased by 46 ± 25%. On the other hand, the fraction of CD34+CD38- cells became 52.0 ± 29% of all CD34+ cells. The absolute number of CD34+CD38- cells was expanded on average 15 ± 12-fold when CD34+ cells were cultured with TPO + FL + KL for 7 days. The expression of CD62L, HLA-DR and CD117 was modulated after culture, particularly with TPO + FL + KL, explaining differences between the adhesion and engraftment of primary and cultured candidate stem cells. We conclude that culture of CD34+ cells with TPO + FL + KL results in a significant increase in the number of candidate stem cells with the CD34+CD38- phenotype

show abstract

Humanized mouse model: Hematopoietic stemcell transplantation and tracking using short tandem repeat technology

Walcher

Hilger

Wege

et al. 2020

Immunity Inflam & Disease

View full text Add to dashboard Cite

Introduction: Models of mice carrying a human immune system, so-called humanized mice, are used increasingly as preclinical models to bridge the gap between model organisms and human beings. Challenges of the humanized mouse model include finding suitable sources for human hematopoietic stem cells (HSC) and reaching sufficient engraftment of these cells in immunocompromised mice. Methods: In this study, we compared the use of CD34 + HSC from cord blood (CB) vs HSC from adult mobilized peripheral blood. Furthermore, we developed a simple and highly specific test for donor identification in humanized mice by applying the detection method of short tandem repeats (STR). Results: It was found that, in vitro, CB-derived and adult HSC show comparable purity, viability, and differentiation potential in colonyforming unit assays. However, in vivo, CB-derived HSC engrafted to a significantly higher extent in NOD.Cg-Prkdc scid IL2rγ tm1Wjl /SzJ (NSG) mice than adult HSC. Increasing the cell dose of adult HSC or using fresh cells without cryopreservation did not improve the engraftment rate. Interestingly, when using adult HSC, the percentage of human cells in the bone marrow was significantly higher than that in the peripheral blood. Using the STR-based test, we were able to identify and distinguish human cells from different donors in humanized mice and in a humanized allogeneic transplantation model.

show abstract

Phenotypic Characterization of Immunomagnetically Purified Umbilical Cord Blood CD34+ Cells

Cited by 22 publications

References 18 publications

Both CD34+38+ and CD34+38− Cells Home Specifically to the Bone Marrow of NOD/LtSZ scid/scid Mice but Show Different Kinetics in Expansion

Both CD34+38+ and CD34+38− Cells Home Specifically to the Bone Marrow of NOD/LtSZ scid/scid Mice but Show Different Kinetics in Expansion

Immunophenotype of hematopoietic stem cells from placental/umbilical cord blood after culture

Humanized mouse model: Hematopoietic stemcell transplantation and tracking using short tandem repeat technology

Contact Info

Product

Resources

About