Extensive phenotypic analysis of CD34 subsets in successive collections of mobilized peripheral blood progenitors

Perey, L; Peters, Rowayda; Pampallona, Sandro; Schneider, Philippe; Groß, Nicole; Leyvraz, Serge

doi:10.1046/j.1365-2141.1998.01062.x

Cited by 17 publications

(9 citation statements)

References 46 publications

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“…Appropriate isotype-matched, nonreactive fluorochrome-conjugated antibodies were used as controls. Cells were stained and analyzed as described previously [22], [23], [24] using a FACS Calibur flow cytometer (BD Biosciences) and data were analyzed using CellQuest Pro software.…”

Section: Methodsmentioning

confidence: 99%

Efficient Generation of Multipotent Mesenchymal Stem Cells from Umbilical Cord Blood in Stroma-Free Liquid Culture

et al. 2010

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BackgroundHaematopoiesis is sustained by haematopoietic (HSC) and mesenchymal stem cells (MSC). HSC are the precursors for blood cells, whereas marrow, stroma, bone, cartilage, muscle and connective tissues derive from MSC. The generation of MSC from umbilical cord blood (UCB) is possible, but with low and unpredictable success. Here we describe a novel, robust stroma-free dual cell culture system for long-term expansion of primitive UCB-derived MSC.Methods and FindingsUCB-derived mononuclear cells (MNC) or selected CD34+ cells were grown in liquid culture in the presence of serum and cytokines. Out of 32 different culture conditions that have been tested for the efficient expansion of HSC, we identified one condition (DMEM, pooled human AB serum, Flt-3 ligand, SCF, MGDF and IL-6; further denoted as D7) which, besides supporting HSC expansion, successfully enabled long-term expansion of stromal/MSC from 8 out of 8 UCB units (5 MNC-derived and 3 CD34+ selected cells). Expanded MSC displayed a fibroblast-like morphology, expressed several stromal/MSC-related antigens (CD105, CD73, CD29, CD44, CD133 and Nestin) but were negative for haematopoietic cell markers (CD45, CD34 and CD14). MSC stemness phenotype and their differentiation capacity in vitro before and after high dilution were preserved throughout long-term culture. Even at passage 24 cells remained Nestin+, CD133+ and >95% were positive for CD105, CD73, CD29 and CD44 with the capacity to differentiate into mesodermal lineages. Similarly we show that UCB derived MSC express pluripotency stem cell markers despite differences in cell confluency and culture passages.Further, we generated MSC from peripheral blood (PB) MNC of 8 healthy volunteers. In all cases, the resulting MSC expressed MSC-related antigens and showed the capacity to form CFU-F colonies.ConclusionsThis novel stroma-free liquid culture overcomes the existing limitation in obtaining MSC from UCB and PB enabling so far unmet therapeutic applications, which might substantially affect clinical practice.

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Section: Methodsmentioning

confidence: 99%

Efficient Generation of Multipotent Mesenchymal Stem Cells from Umbilical Cord Blood in Stroma-Free Liquid Culture

et al. 2010

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“…First, we did not assess the subpopulation of CD34+ cells. It was demonstrated that a majority of CD34+ cells expressed CD13 and CD33, which represent myeloid marker [27]. Erythroid, megakaryocytic lineage and EPC occupy only a small portion of CD34+ cells.…”

Section: Discussionmentioning

confidence: 99%

rHuEPO Dose Inversely Correlated with the Number of Circulating CD34+ Cells in Maintenance Hemodialysis Patients

Kohagura¹,

Ohya²,

Miyagi³

et al. 2007

Nephron Clin Pract

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Background: A decreased number of endothelial progenitor cells (EPCs) as well as anemia have been reported to be associated with cardiovascular disease. Maintenance hemodialysis (MHD) patients who require higher doses of recombinant human erythropoietin (rHuEPO) have higher cardiovascular mortality. However, it has not been examined whether there is correlation between the numbers of CD34+ cells, including EPCs and erythroid progenitor cells, and the dose of rHuEPO in MHD patients. Methods: We measured the number of circulating CD34+ cells by flow cytometry and examined the clinical characteristics in 35 MHD patients (50% male). Results: A significant negative correlation was discovered between the number of circulating CD34+ cells and the dose of rHuEPO (r = –0.441, p = 0.013). We performed multivariate regression analysis to determine whether the number of CD34+ cells was associated with age, gender, diabetes, serum albumin, C-reactive protein, ferritin, statin, and dose of rHuEPO. The dose of rHuEPO, diabetes, and statin were independent predictors of the number of circulating CD34+ cells. A reciprocal analysis that divided these patients into two groups according to mean value of CD34+ cells also demonstrated the significant relationship between rHuEPO dose level and the number of CD34+ cells. Conclusion: These findings suggested that the requirement of a higher dose of rHuEPO to maintain target hemoglobin was associated with a decrease in the number of CD34+ cells. This relationship may be partly responsible for the higher cardiovascular mortality of this group among MHD patients.

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“…In normal and neoplastic hematopoietic populations, CD13 is expressed specifically on the earliest cell committed to the myeloid lineage (CFU-GM) and all of its differentiated progeny and is a reliable diagnostic marker for certain types of leukemia [3]. More recently, CD13 has been shown to be highly expressed on adult pluripotent stem cells derived from multiple sites [15][16][17] and differentiated ES cells [18,19], as well as subsets of peripheral blood progenitor cells [20]. In addition, CD13 has been postulated to play a role in fetal liver hematopoiesis [21] and myeloid cell differentiation [22].…”

Section: The Hematopoietic Profile Of Cd13 Null Mice Is Normalmentioning

confidence: 99%

CD13 is dispensable for normal hematopoiesis and myeloid cell functions in the mouse

Winnicka

O'Conor

Schacke

et al. 2010

Journal of Leukocyte Biology

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The robust and consistent expression of the CD13 cell surface marker on very early as well as differentiated myeloid hematopoietic cells has prompted numerous investigations seeking to define roles for CD13 in myeloid cells. To address the function of myeloid CD13 directly, we created a CD13 null mouse and assessed the responses of purified primary macrophages or DCs from WT and CD13 null animals in cell assays and inflammatory disease models, where CD13 has been implicated previously. We find that mice lacking CD13 develop normally with normal hematopoietic profiles except for an increase in thymic but not peripheral T cell numbers. Moreover, in in vitro assays, CD13 appears to be largely dispensable for the aspects of phagocytosis, proliferation, and antigen presentation that we tested, although we observed a slight decrease in actin-independent erythrocyte uptake. However, in agreement with our published studies, we show that lack of monocytic CD13 completely ablates anti-CD13-dependent monocyte adhesion to WT endothelial cells. In vivo assessment of four inflammatory disease models showed that lack of CD13 has little effect on disease onset or progression. Nominal alterations in gene expression levels between CD13 WT and null macrophages argue against compensatory mechanisms. Therefore, although CD13 is highly expressed on myeloid cells and is a reliable marker of the myeloid lineage of normal and leukemic cells, it is not a critical regulator of hematopoietic development, hemostasis, or myeloid cell function.

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Extensive phenotypic analysis of CD34 subsets in successive collections of mobilized peripheral blood progenitors

Cited by 17 publications

References 46 publications

Efficient Generation of Multipotent Mesenchymal Stem Cells from Umbilical Cord Blood in Stroma-Free Liquid Culture

Efficient Generation of Multipotent Mesenchymal Stem Cells from Umbilical Cord Blood in Stroma-Free Liquid Culture

rHuEPO Dose Inversely Correlated with the Number of Circulating CD34+ Cells in Maintenance Hemodialysis Patients

CD13 is dispensable for normal hematopoiesis and myeloid cell functions in the mouse

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