Obtaining of CD34+ cells from healthy blood donors: development of a rapid and efficient procedure using leukoreduction filters

Peytour, Yann; Guitart, Amélie V.; Villacreces, Arnaud; Chevaleyre, J.; Lacombe, Francis; Ivanović, Zoran; Perreten, Vincent

doi:10.1111/j.1537-2995.2010.02683.x

Cited by 16 publications

(8 citation statements)

References 12 publications

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“…Because both in vitro and ex vivo results suggested that autophagy is required for the full maintenance of LC2 repopulation ability of CML cells incubated in hypoxia, we asked whether it also plays a critical role in normal CD34 + cells. Normal CD34 + cells were purified from leukoreduction filters as described Peytour et al [ 20 ]. Four days after infection, normal CD34 + cells were analyzed for GFP expression and between 35 and 50% of GFP + cells were detected in 3 independent experiments (Figure 3A ).…”

Section: Resultsmentioning

confidence: 99%

Chronic myeloid leukemia progenitor cells require autophagy when leaving hypoxia-induced quiescence

et al. 2017

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Albeit tyrosine kinase inhibitors anti-Abl used in Chronic Myeloid Leukemia (CML) block the deregulated activity of the Bcr-Abl tyrosine kinase and induce remission in 90% of patients, they do not eradicate immature hematopoietic compartments of leukemic stem cells. To elucidate if autophagy is important for stem cell survival and/or proliferation, we used culture in low oxygen concentration (0.1% O2 for 7 days) followed back by non-restricted O2 supply (normoxic culture) to mimic stem cell proliferation and commitment. Knockdown of Atg7 expression, a key player in autophagy, in K562 cell line inhibited autophagy compared to control cells. Upon 7 days at 0.1% O2 both K562 and K562 shATG7 cells stopped to proliferate and a similar amount of viable cells remained. Back to non-restricted O2 supply K562 cells proliferate whereas K562 shATG7 cells exhibited strong apoptosis. Using immunomagnetic sorted normal and CML CD34+ cells, we inhibited the autophagic process by lentiviral infection expressing shATG7 or using a Vps34 inhibitor. Both, normal and CML CD34+ cells either competent or deficient for autophagy stopped to proliferate in hypoxia. Surprisingly, while normal CD34+ cells proliferate back to non restricted O2 supply, the CML CD34+ cells deficient for autophagy failed to proliferate. All together, these results suggest that autophagy is required for CML CD34+ commitment while it is dispensable for normal CD34 cells.

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Section: Resultsmentioning

confidence: 99%

Chronic myeloid leukemia progenitor cells require autophagy when leaving hypoxia-induced quiescence

et al. 2017

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“…These cells are still underutilized in research and medicine most likely due to their low frequency in blood, despite the fact that they are easily accessible without any invasive procedures nor side effects for donors. Moreover our group has shown in previous studies that the use of LRF from different blood donors allows us to obtain enough hematopoietic stem and progenitor cells that is CD34 + cells for research studies (Peytour et al, ; Peytour, Villacreces, Chevaleyre, Ivanovic, & Praloran, ). In the present work, we evidenced some characteristics and functional properties of SSPB HSC.…”

Section: Discussionmentioning

confidence: 99%

“…These rare and relatively old findings suggest that HSCs from SSPB have an interesting biological potential and might be of great interest for research and medicine, as they also represent a readily accessible and safe source of HSCs. In this context, our group recently showed the possibility of using CD34 + cells from SSPB by recycling cells retained on leuko‐reduction filters (LRF) from whole blood samples (Peytour et al, ). We proved that these CD34 + cells contain HSCs with short‐ and long‐term engraftment capacity (Brunet et al, ).…”

Section: Introductionmentioning

confidence: 99%

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

Bourdieu

Avalon

Lapostolle

et al. 2017

Journal Cellular Physiology

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“…Leukocytes were recovered from leukodepletion filters (T2975, Fresenius Kabi, Louviers, France) by counterflow elution as described elsewhere 6 , 19 , 20 with a slight modification, i.e. the cells were flushed directly into 50 mL tubes (Falcon, Dutscher, Brumath, France) (see Online Supplementary Methods ).…”

Section: Methodsmentioning

confidence: 99%

Repopulating hematopoietic stem cells from steady-state blood before and after ex vivo culture are enriched in the CD34⁺CD133⁺CXCR4^low fraction

et al. 2018

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The feasibility of ex vivo expansion allows us to consider the steady-state peripheral blood as an alternative source of hematopoietic stem progenitor cells for transplantation when growth factor-induced cell mobilization is contraindicated or inapplicable. Ex vivo expansion dramatically enhances the in vivo reconstituting cell population from steady-state blood. In order to investigate phenotype and the expression of homing molecules, the expression of CD34, CD133, CD90, CD45RA, CD26 and CD9 was determined on sorted CD34+ cells according to CXCR4 (“neg”, “low” “bright”) and CD133 expression before and after ex vivo expansion. Hematopoietic stem cell activity was determined in vivo on the basis of hematopoietic repopulation of primary and secondary recipients - NSG immuno-deficient mice. In vivo reconstituting cells in the steady-state blood CD34+ cell fraction before expansion belong to the CD133+ population and are CXCR4low or, to a lesser extent, CXCR4neg, while after ex vivo expansion they are contained only in the CD133+CXCR4low cells. The failure of the CXCR4bright population to engraft is probably due to the exclusive expression of CD26 by these cells. The limiting-dilution analysis showed that both repopulating cell number and individual proliferative capacity were enhanced by ex vivo expansion. Thus, steady-state peripheral blood cells exhibit a different phenotype compared to mobilized and cord blood cells, as well as to those issued from the bone marrow. These data represent the first phenotypic characterization of steady-state blood cells exhibiting short- and long-term hematopoietic reconstituting potential, which can be expanded ex vivo, a sine qua non for their subsequent use for transplantation.

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Obtaining of CD34+ cells from healthy blood donors: development of a rapid and efficient procedure using leukoreduction filters

Cited by 16 publications

References 12 publications

Chronic myeloid leukemia progenitor cells require autophagy when leaving hypoxia-induced quiescence

Chronic myeloid leukemia progenitor cells require autophagy when leaving hypoxia-induced quiescence

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

Repopulating hematopoietic stem cells from steady-state blood before and after ex vivo culture are enriched in the CD34⁺CD133⁺CXCR4^low fraction

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Obtaining of CD34+ cells from healthy blood donors: development of a rapid and efficient procedure using leukoreduction filters

Cited by 16 publications

References 12 publications

Chronic myeloid leukemia progenitor cells require autophagy when leaving hypoxia-induced quiescence

Chronic myeloid leukemia progenitor cells require autophagy when leaving hypoxia-induced quiescence

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

Repopulating hematopoietic stem cells from steady-state blood before and after ex vivo culture are enriched in the CD34+CD133+CXCR4low fraction

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Repopulating hematopoietic stem cells from steady-state blood before and after ex vivo culture are enriched in the CD34⁺CD133⁺CXCR4^low fraction