A-S Espadinha et al, Oncotarget 2017

A tyrosine kinase-STAT5-miR21-PDCD4 regulatory axis in chronic and acute myeloid leukemia cells


MicroRNAs (miRNAs) are regulators of several key patho-physiological processes, including cell cycle and apoptosis. Using microarray-based miRNA profiling in K562 cells, a model of chronic myeloid leukemia (CML), we found that the oncoprotein BCR-ABL1 regulates the expression of miR-21, an “onco-microRNA”, found to be overexpressed in several cancers. This effect relies on the presence of two STAT binding sites on the promoter of miR-21, and on the phosphorylation status of STAT5, a transcription factor activated by the kinase activity of BCR-ABL1. Mir-21 regulates the expression of PDCD4 (programmed cell death protein 4), a tumor suppressor identified through a proteomics approach. The phosphoSTAT5 — miR-21 — PDCD4 pathway was active in CML primary CD34+ cells, but also in acute myeloid leukemia (AML) models like MV4.11 and MOLM13, where the constitutively active tyrosine kinase FLT3-ITD plays a similar role to BCR-ABL1 in the K562 cell line.


(STEM ALPHA.A #5510)

C.S. Jackson et al. / Stem Cell Research

Targeting the aryl hydrocarbon receptor nuclear translocator complexwith DMOG and Stemregenin 1 improves primitive hematopoietic stemcell expansion


Culture conditions used for the expansion of hematopoietic stem and progenitor cells (HSCs and HPCs, collective-ly HSPCs) should ideally favor the self renewal of long-term HSCs. At 20% O2, the synthesis of HIF-1αis balancedby its hydroxylation and proteasomal degradation. This favors HSPC differentiation, but can be prevented by cul-turing CD34+ cord blood cells in the presence of dimethyloxaloylglycine (DMOG). This differentiation may alsobe reduced by culturingthe cells in the presence of Stemregenin 1,anantagonistof the aryl hydrocarbon receptor(AhR). The objective of this study was to investigate how hypoxia, DMOG and Stemregenin 1 might affect the ex-pansion of HSPCs with the aim of identifying optimalconditions for expansionin culture. It was foundthatDMOGdecreased proliferation but was effective in preserving the number of cells in the primitive hematopoietic sub-populationsin vitro. The effect of DMOG was similar to hypoxia, although differences were observed with regardto the side population and CD34+ sub-populations. Stemregenin 1 on the other hand increased the size of theprimitive as well as the other HSC sub-populations. The use of Stemregenin 1 with DMOG increased the propor-tion ofprimitive HSCs to 3.54% comparedto2.61% for Stemregenin 1 alone.Invivoengraftment studiesconfirmedthesefindings and showed that fewer cells (3710) are required for long-term engraftment when HSCs are grownin Stemregenin 1 together with hypoxia than in Stemregenin 1 under conditions of normoxia (13430).


(STEM ALPHA.1D #5112)

A Bourdieu et al, J Cell Physiol 2017

Steady state peripheral blood provides cells with functional and metabolic characteristics of real hematopoietic stem cells.

J Cell Physiol.  2017;  (ISSN: 1097-4652)

Bourdieu A; Avalon M; Lapostolle V; Ismail S; Mombled M; Debeissat C; Guérinet M; Duchez P; Chevaleyre J; Vlaski-Lafarge M; Villacreces A; Praloran V; Ivanovic Z; Brunet de la Grange P


Hematopoietic stem cells (HSCs), which are located in the bone marrow, also circulate in cord and peripheral blood. Despite high availability, HSCs from steady state peripheral blood (SSPB) are little known and not used for research or cell therapy. We thus aimed to characterize and select HSCs from SSPB by a direct approach with a view to delineating their main functional and metabolic properties and the mechanisms responsible for their maintenance. We chose to work on Side Population (SP) cells which are highly enriched in HSCs in mouse, human bone marrow, and cord blood. However, no SP cells from SSBP have as yet been characterized. Here we showed that SP cells from SSPB exhibited a higher proliferative capacity and generated more clonogenic progenitors than non-SP cells in vitro. Furthermore, xenotransplantation studies on immunodeficient mice demonstrated that SP cells are up to 45 times more enriched in cells with engraftment capacity than non-SP cells. From a cell regulation point of view, we showed that SP activity depended on O2 concentrations close to those found in HSC niches, an effect which is dependent on both hypoxia-induced factors HIF-1α and HIF-2α. Moreover SP cells displayed a reduced mitochondrial mass and, in particular, a lower mitochondrial activity compared to non-SP cells, while they exhibited a similar level of glucose incorporation. These results provided evidence that SP cells from SSPB displayed properties of very primitive cells and HSC, thus rendering them an interesting model for research and cell therapy.  


Product used : STEM LAPHA.1D (#5112)

S Ducassou, T.Journal of Pathology 2017

MYB–GATA1 fusion promotes basophilic leukaemia: involvement of interleukin-33 and nerve growth factor receptors



Acute basophilic leukaemia (ABL) is a rare subtype of acute myeloblastic leukaemia. We previously described a recurrent t(X;6)(p11;q23) translocation generating an MYB–GATA1 fusion gene in male infants with ABL. To better understand its role, the chimeric MYB–GATA1 transcription factor was expressed in CD34-positive haematopoietic progenitors, which were transplanted into immunodeficient mice. Cells expressing MYB–GATA1 showed increased expression of markers of immaturity (CD34), of granulocytic lineage (CD33 and CD117), and of basophilic differentiation (CD203c and FcϵRI). UT-7 cells also showed basophilic differentiation after MYB–GATA1 transfection. A transcriptomic study identified nine genes deregulated by both MYB–GATA1 and basophilic differentiation. Induction of three of these genes (CCL23, IL1RL1, and NTRK1) was confirmed in MYB–GATA1-expressing CD34-positive cells by reverse transcription quantitative polymerase chain reaction. Interleukin (IL)-33 and nerve growth factor (NGF), the ligands of IL-1 receptor-like 1 (IL1RL1) and neurotrophic receptor tyrosine kinase 1 (NTRK1), respectively, enhanced the basophilic differentiation of MYB–GATA1-expressing UT-7 cells, thus demonstrating the importance of this pathway in the basophilic differentiation of leukaemic cells and CD34-positive primary cells. Finally, gene reporter assays confirmed that MYB and MYB–GATA1 directly activated NTRK1 and IL1RL1 transcription, leading to basophilic skewing of the blasts. MYB–GATA1 is more efficient than MYB, because of better stability. Our results highlight the role of IL-33 and NGF receptors in the basophilic differentiation of normal and leukaemic cells. Copyright © 2017 Pathological Society of Great Britain and Ireland. Published by John Wiley & Sons, Ltd.



(STEM ALPHA.1D #5112)

Contrôle Progéniteurs Hématopoïétiques

BILAN 2009 - 2010 ET ÉVOLUTION DU CONTRÔLE EXTERNE DES CELLULES SOUCHES HEMATOPOÏETIQUES B. Panterne, C. Sabatini, V. Duval, G. Huyghe*, L. Fleury et L. Mouillot Agence francaise de sécurite sanitaire des produits de santé (Afssaps) – Direction des laboratoires et des controles (DLC) – Unité Produits sanguins et therapie cellulaire. *Unite Microbiologie, 143/147, Bd Anatole France, 93285 Saint-Denis Cedex


B.Panterne et al, 2010. Contrôle de qualité des greffons de sang placentaire décongelés : résultats d’une étude multicentrique. Transfusion clinique et Biologique 17 (2010) 41-46


Ten Years of External Quality Control for Cellular Therapy Products in France

Béatrice Panterne et al.*Direction des Laboratoires et des Contrôles, Agence Française de Sécurité Sanitaire des Produits de Santé (Afssaps) France


Delessalle N. et al, 2007. Contrôle de qualité externe des milieux utilisés dans la greffe de cornée : résultats des tours de contrôles de 2005 à 2006. AFSSAPS - DLC, unité Produits sanguins et thérapie cellulaire St Denis et Microbiologie.

Panterne B, et al, 2007. Le test clonogénique CFU-GM dans le contexte du contrôle externe des CSH, AFSSAPS - DLC St Denis.


PANTERNE B, et al. Bilan 2003 - 2004 du contrôle externe des cellules souches hématopoïétiques. Cas des Moelles osseuses et des CSP décongelées, SFTS 2005, poster AFSSAPS.

Sabatini C., Panterne B., MouilloT L (2000). Comparaison de milieux semi-solides prêts à l’emploi pour le test clonogénique des progéniteurs CFU-GM. XXème Congrès de la Société Française de Transfusion Sanguine. Paris le 27-29 Juin 2000.Transfusion Clinique et Biologique vol.7 n°3 P314.

Projet de recommandation (power point) en octobre 2010




Dobo I, Pineau D, Robillard N, Genevieve F, Piard N, Zandecki M, Hermouet S. Standardization of the CFU-GM Assay: Advantages of Plating a Fixed Number of CD34(+) Cells in Collagen Gels. J Hematother Stem Cell Res. (2003) Oct ; 12 (5) : 543-51.


Milieu liquide sans sérum STEM ALPHA.A

En 2012

Mohammad Hammoud et al. 2012. Combination of low O2 concentration and mesenchymal stromal cells during culture of cord blood CD34+ cells improves the maintenance and proliferative capacity of hematopoietic stem cells. 

Article first published online: 24 FEB 2012 DOI: 10.1002/jcp.23019

Journal of Cellular Physiology Vol 227, Issue 6, p. 2750–2758, June 2012


En 2011

Membrane-bound IL-15 stimulation on peripheral blood NK ...

de S Negrini - 2011 - Published Ahead of Print on January 17, 2011,  expanded at high cell density in STEMα−Α medium (Stem Alpha, Saint Clement les places, ...)


En 2010

Modeling of congenital erythropoietic porphyria by RNA interference: a new tool for preclinical gene therapy evaluation

Elodie Robert-Richard1,2, Magalie Lalanne1,2, Isabelle Lamrissi-Garcia1,2, Véronique Guyonnet-Duperat1, Emmanuel Richard1,2, Vincent Pitard2,3, Fréderic Mazurier1,2, François Moreau-Gaudry1,2, Cécile Ged1,2, Hubert de Verneuil1,2,*

The Journal of Gene Medicine

Volume 12, Issue 8, pages 637–646, August 2010


En 2009

Low O2 concentrations enhance the positive effect of IL-17 on the maintenance of erythroid progenitors during co-culture of CD34+ and mesenchymal stem cells

Eur. Cytokine Netw., Vol. 20 n° 1, March 2009, 10-6


Aleksandra Krstić1, Marija Vlaski2,3, Mohammad Hammoud2, Jean Chevaleyre2,

Pascale Duchez2, Gordana Jovčić1, Diana Bugarski1, Pavle Milenković1, Philippe Bourin4,Jean-Michel Boiron2,3, Vincent Praloran3,5, Zoran Ivanović2

1 Institute for Medical Research, Belgrade University, Belgrade, Serbia

2 French Blood Institute, Aquitaine-Limousin Branch, 5, place Amélie Raba Léon, BP 24, 33035 Bordeaux Cedex, France

3 Victor-Ségalen University (Bordeaux 2), Bordeaux, France

4 French Blood Institute, Pyrénées-Méditerranée Branch, Toulouse, France

5 CNRS UMR 5164 Correspondence: Z. Ivanovic, MD, PhD, Etablissement Français du Sang Aquitaine-Limousin, 5 place Amélie Raba Léon, BP 24,

33035 Bordeaux Cedex, France <zoran.ivanovic@efs.sante.fr> Accepted for publication January 16, 2009


Tissue inhibitors of matrix metalloproteinases in platelets and megakaryocytes: A novel organization for these secreted proteins

Julien Villeneuve,  Anna Block Marie-Caroline Le Bousse-Kerdilès, Sébastien Lepreux Paquita Nurden

Jean Ripoche  Alan T. Nurden

Experimental Hematology Volume 37, Issue 7 , Pages 849-856, July 2009


Review of Althought - Definition and Scientific Information - Page 1

... detected was similar in all media tested: 29 +- 10% in Stem alpha-A, 28 +- 9,7% in Stemalpha-AMK, 28 +-7,7% in Stemalpha-AMK+TPO For normal subjects, ... eurekamag.com/keyword/a/187/althought.php - États-Unis


Giuliani M, et al., 2008. Generation of a Novel Regulatory NK Cell Subset from Peripheral Blood CD34+ Progenitors Promoted by Membrane-Bound IL-15. PloS ONE (2008), 3, Issue 5, e2241, 1-16.

Hermitte F., Brunet de la Grange P., Belloc F., Praloran V. and Ivanovic Z. (2006) Very low O2 concentration (0.1%) favors G0 return dividing CD34+ cells. Stem Cells, 24, 1, 2006, 65-73.

Giron-Michel J., .... and Azzarone B. Membrane bound and soluble IL-15/IL-15Ra complexes display differential signaling and functions on human hematopoietic progenitors. Blood, 1 october 2005, 106, 7, 2302-2310


Ivanovic Z., 2004. Interleukine-3 and ex vivo maintenance of hematopoietic stem cells : facts and controversies. Eur. Cytokine Netw, 15, 6-13.

Ivanovic Z, Hermitte F, Brunet de la Grange P, Dazey B, Belloc F, Lacombe F, Vezon G and Praloran V. (2004) Simultaneous maintenance of human cord blood SCID-repopulating cells and expansion of commited progenitors at low O2 concentration (3%). Stem Cell 03 -0131.R1.

Desplat V., Faucher J-L., Mahon F-X., Dello Sbarba P., Praloran V. and Ivanovic Z. (2002). Hypoxia modifies proliferation and differentiation of CD34+ CML cells, STEM CELLS, 20 : 347-354.

De Bruyn C., et al (2003). Ex vivo myeloid differenciation of cord blood CD34+ cells : comparison of four sreum-free media containing bovine or human albumin, Cytotherapy, 5, no 2, 153-160 .

De Bruyn C., et al (2003). Ex vivo expansion of neutrophil precursor cells from fresh and cryopreserved cord blood cells, Cytotherapy, 5, no 1, 87-98.

Tanja Rossmanith, 2001. Kultivierung von CD34+ -Zellen aus NabelSchnurblut zur ex vivo expansion von Stamm-und Vorlauferzellen und unterschungen zu deren homing-fahigkeiten.

Abnese P., Leboeuf M., Rosa J-P. and Uzan G. (2000). Identification of GATA-overlaping sequence within the enhancer of the murine GPIIb promoter that induces transcriptional deregulation in human K562 cells, Blood, 96, 4, 1348-1357.

Lepage A., Leboeuf M., Cazenave J-P., De la salle C., Lanza F. and Uzan G. (2000). The aIIIbb3 integrin and GPIb-V-IX complex identify distinct stages in the maturation of CD34+ cord blood cells to megacaryocytes, Blood, 96, 13, 4169-4177.


Cell.souches/Progéniteurs : Leucémies

The peripheral CD138+ population but not the CD138) population contains myeloma clonogenic cells in plasma cell leukaemia patients 2011 Blackwell Publishing Ltd, British Journal of Haematology doi:10.1111/j.1365-2141.2011.08904.x David Chiron, Sylvanie Surget, Sophie Maı¨ga, Re´gis Bataille, , Philippe Moreau, , Steven Le Gouil, Martine Amiot,  , Catherine Pellat-Deceunynck


Leukemia 24, 601-612 (March 2010)

Functional characterization of high levels of meningioma 1 as collaborating oncogene in acute leukemia

T Liu, D Jankovic, L Brault, S Ehret, F Baty, V Stavropoulou, V Rossi, A Biondi and J Schwaller



Belloc F., et al, 2009. The stem cell factor c KIT pathway must be inhibited to enable apoptosis induced by BRC-ABL inhibitors in chronic myelogenous leukemia cells. Leukemia, 2009, 1-7.

Pigneux A., et al, 2008. Triptolide cooperates with chemotherapy to induce apoptosis in acute myeloid leukemia cells. Experimental Hematology, EXPHEM2374_proof _ 16-9-2008 0.

Emmanuelle Me´noret, et al, 2008. IL-21 Stimulates Human Myeloma Cell Growth through an Autocrine IGF-1 Loop. The Journal of Immunology, 6837-6842.

Smadja D., et al, (2008). Bone Morphogenetic Proteins 2 and 4 Are Selectively Expressed by Late Outgrowth Endothelial Progenitor Cells and Promote Neoangiogenesis, Arterioscler. Thromb. Vasc. Biol.., 2008, 1-7.

Chabanon A., et al, 2008. A CROSS-TALK between SDF-1 and TGF-β CONTROLS THE QUIESCENCE/CYCLING SWITCH OF CD34+ PROGENITORS THROUGH FoxO3 AND mTOR. Stem Cells, 2008; doi:10.1634/stemcells.2008-0219.

James C., et al., 2008. The hematopoietic stem cell compartment ok JAK2V617F positive myeloproliferative disorders is a reflection of disease heterogeneity. Blood, (2008), 112, n°6, 2429-2438.

Tiedt R, et al, 2008. Ratio of mutant JAK2-V617F of wild-type Jak2 determines the MPD phenotypes in transgenic mice. Blood (2008), 111 : 3931-3940.

Larghero J, et al, 2008. Phenotypical and functional characteristics of in vitro expanded bone marrow mesenchymal stem cells from patients with systemic sclerosis. Ann Rheum Dis (2008), 67 : 443-449.

Colette M., et al (2007). Crucial role of phosphatase CD45 in determining signalling and proliferation of human myeloma cells, Eur cytokine Netw., vol 18 n°3, 2007, 1-7.

Arnulf B. et al. (2007) Phenotypic and functional characterization of bone marrow mesenchymal stem cells derived from patients with multiple myeloma. Leukemia, (2007), 21, 158-163.

Robert-Richard E., Ged C., Ortet J., Santarelli X., Lamrissi-Garcia I., De Verneuil H., Mazurier F. (2006). Human cell engraftment after busulfan or irradiation conditioning of NOD/SCID mice. Haematologica 2006, 91 : 1384-1387.

Dobo I., et al. (2006). Abnormal peripheral blood progenitors are consistently observed after cell culture in myelodysplasic syndrome (MDS), allowing the diagnosis of eraly step of the disorder. Haematologica, 2006, 91 (s1-0945), 11è Congress of the European Hematology Association.

Girodon F. et al. Is ageing associated with a decrease of BFU-E or CFU-GM growth pattern studied by in vitro cultures ?

Isabelle Corre-Buscail, Danielle Pineau, Marjorie Boissinot and Sylvie Hermouet , 2005. Erythropoietin-independent erythroid colony formation by bone marrow progenitors exposed to interleukin-11 and interleukin-8. Experimental Hematology, Volume 33, Issue 11, November 2005, Pages 1299-1308.

Lataillade JJ., et al., 2005. Phenotypic and functional characteristics of CD34+ cells are related to their anatomical environment : is their versatility a prerequisite for their bio-availability ? Journal of Leukocyte Biology, 77 : 634-643.

J. van Grevenynghe et al. (2005) Human CD34-positive hematopoietic stem cells constitue targets for carcinogenic polycyclic aromatic hydrocarbons. JPET 314 : 693-702.

Atfi A, Abécassis L and Bourgeade MF. Brc-Abl activates the AKT/FoxO3 signalling pathway to restrict transforming growth factor-b-mediated cytostatic signals. EMBO reports AOP, 19 august 2005.

Boudard D., Viallet A., Piselli S., Guyotat D. and Campos L. (2003) In vitro study of stromal cell defects in myelodysplastic syndromes, Haematologica, 88 : 827-829.

Stephane Prost, Magali LeDiscorde, Rima Haddad, Jean-Claude Gluckman, Bruno Canque and Marek Kirszenbaum, 2002. Characterization of a Novel Hematopoietic Marker Expressed from Early Embryonic Hematopoietic Stem Cells to Adult Mature Lineages, Blood Cells, Molecules, and Diseases, Volume 29, Issue 2, September 2002, Pages 236-248.

Lataillade JJ., Clay D., Bourin P., Hérodin F., Dupuy C., Jasmin C. and Le Bousse-Kerdiles MC. (2002). Stromal cell-derived factor 1 regulates primitive hematopiesis by suppressing apoptosis and by promoting G0/G1 transition in CD34+ cells : evidence for an autocrine/paracrine mechanism. Blood, 99, 4, 1117-1129.

Brunet de la Grange P., Ivanovic Z., Leprivey-Lorgeot V. and Praloran V. (2002). Angiotensin II that reduces the colony-forming ability of hematopoietic progenitors in serum free medium has an inverse effect in serum-supplemented medium, Stem Cells, 20 : 269-271.

Boudard D, Viallet A, Piselli S, Licari D, Guyotat D and Campos L. (2002) A multiparametric study of bone marrow stromal cells in myelodisplasic syndromes (article accepté dans Leukemia).

Dobo I., et al. (2001). Comparison of four serum-free, cytokine-free media for analysis of endogenous erythroid colony growth in polycythemia vera and essential thrombocytemia. The hematology journal, 2, 396-403.

Lataillade J-J., et al. (2000). Chemokine SDF-1 enhances circulating CD34+ cell proliferation in synergy with cytokines : possible role in progenitor survival, Blood, 95, 756-768.