Phenotyping of peripheral blood hemopoietic progenitor cells - in vitro cultures using CD34-/CD33-immunomagnetic purging

2009 ◽  
Vol 47 (5) ◽  
pp. 361-366 ◽  
Author(s):  
Stefan Serke ◽  
Yoshikazu Abe ◽  
Andreas Kirsch ◽  
Dieter Huhn
1995 ◽  
Vol 16 (5-6) ◽  
pp. 483-492 ◽  
Author(s):  
Juergen Thiele ◽  
Claudia Wickenhauser ◽  
Stephan E. Baldus ◽  
Thomas Kuemmel ◽  
Thomas K. Zirbes ◽  
...  

1995 ◽  
Vol 93 (2-4) ◽  
pp. 83-90 ◽  
Author(s):  
Claudia Wickenhauser ◽  
Jürgen Thiele ◽  
Uta Drebber ◽  
Hans Michael Kvasnicka ◽  
Andreas Thiel ◽  
...  

Blood ◽  
1985 ◽  
Vol 65 (4) ◽  
pp. 869-876 ◽  
Author(s):  
RM Crapper ◽  
JW Schrader

Abstract Antisera were raised by immunizing rabbits with cloned lines of murine hemopoietic progenitor cells (P cells) that depended on the presence of a specific hemopoietic growth factor, persisting cell-stimulating factor (PSF), for their growth and survival. The unabsorbed antiserum was inhibitory, but after absorption with murine spleen cells and the mastocytoma, P815, significant stimulation of both P cell growth and thymidine incorporation was evident. IgG antibodies isolated from the antiserum by staphylococcal protein A chromatography or further purified by diethylaminoethyl anion exchange chromatography, ammonium sulphate precipitation, and gel filtration using Sephacryl S-300 were responsible for the stimulation. The absorbed antiserum promoted the survival of normal murine bone marrow cells in liquid culture over a four-day period, and the inclusion of IgG antibodies in agar cultures of normal bone marrow promoted the in vitro survival, over a 48-hour period, of cells capable of subsequently generating, in the presence of a source of PSF, colonies of neutrophils, macrophages, and megakaryocytes. It is postulated that the antibodies act by stimulating the PSF receptor on both the factor-dependent cell lines and normal myeloid progenitor cells.


2009 ◽  
Vol 37 (2) ◽  
pp. 170-174 ◽  
Author(s):  
Gemma Molyneux ◽  
Sian Rizzo ◽  
John Turton ◽  
Parvinder Phul ◽  
Frances Gibson

Blood ◽  
1981 ◽  
Vol 58 (2) ◽  
pp. 376-386 ◽  
Author(s):  
NA Nicola ◽  
D Metcalf ◽  
H von Melchner ◽  
AW Burgess

Abstract Hemopoietic progenitor cells (colony- and cluster-forming cells in semisolid agar) were purified from light density CBA murine fetal liver cells using fluorescein-conjugated pokeweed mitogen (PWM) and a rhodamine-conjugated antineutrophil serum sandwich (alpha N) and three- parameter fluorescence-activated cell sorting. All clonable progenitor cells were highly enriched (36–50-fold) in PWM-positive (greater than channel 15), alpha N-negative (less than channel 30) fractions with relatively high intensity (greater than 100) low angle light scatter. No separation was achieved between different types of progenitor cells (granulocyte-macrophage and erythroid colony-forming cells). The enriched fraction was a pure population of large, basophilic, undifferentiated blast cells, and in agar cultures stimulated with colony-stimulating factors, up to 90% of the enriched cells were hemopoietic progenitor cells capable of varying levels of clonal proliferation. Further fractionation based on increasing fluorescence with PWM separated into discrete populations, nonproliferative morphologically recognizable erythroid cells, late erythroid progenitor cells (day 2 CFU-E), and cells forming pure or mixed erythroid burst colonies. In addition, the majority of pluripotential hemopoietic stem cells (CFU-SS) were clearly separated from progenitor cells forming colonies in vitro. The present techniques provide suitable numbers of enriched progenitor cells for a variety of biological and biochemical studies.


2004 ◽  
Vol 24 (2) ◽  
pp. 741-756 ◽  
Author(s):  
Ludovica Bruno ◽  
Reinhard Hoffmann ◽  
Fraser McBlane ◽  
John Brown ◽  
Rajeev Gupta ◽  
...  

ABSTRACT The molecular mechanisms governing self-renewal, differentiation, and lineage specification remain unknown. Transcriptional profiling is likely to provide insight into these processes but, as yet, has been confined to “static” molecular profiles of stem and progenitors cells. We now provide a comprehensive, statistically robust, and “dynamic” analysis of multipotent hemopoietic progenitor cells undergoing self-renewal in response to interleukin-3 (IL-3) and multilineage differentiation in response to lineage-affiliated cytokines. Cells undergoing IL-3-dependent proliferative self-renewal displayed striking complexity, including expression of genes associated with different lineage programs, suggesting a highly responsive compartment poised to rapidly execute intrinsically or extrinsically initiated cell fate decisions. A remarkable general feature of early differentiation was a resolution of complexity through the downregulation of gene expression. Although effector genes characteristic of mature cells were upregulated late, coincident with morphological changes, lineage-specific changes in gene expression were observed prior to this, identifying genes which may provide early harbingers of unilineage commitment. Of particular interest were genes that displayed differential behavior irrespective of the lineage elaborated, many of which were rapidly downregulated within 4 to 8 h after exposure to a differentiation cue. These are likely to include genes important in self-renewal, the maintenance of multipotentiality, or the negative regulation of differentiation per se.


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