2017
DOI: 10.1002/cyto.a.23148
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Multiparametric Whole Blood Dissection: A one‐shot comprehensive picture of the human hematopoietic system

Abstract: Human hematopoiesis is a complex and dynamic system where morphologically and functionally diverse mature cell types are generated and maintained throughout life by bone marrow (BM) Hematopoietic Stem/Progenitor Cells (HSPC). Congenital and acquired hematopoietic disorders are often diagnosed through the detection of aberrant frequency or composition of hematopoietic cell populations. We here describe a novel protocol, called “Whole Blood Dissection” (WBD), capable of analyzing in a single test‐tube, hematopoi… Show more

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Cited by 22 publications
(31 citation statements)
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“…The detailed protocol for whole BM staining has been previously reported (Basso-Ricci et al, 2017). In brief, Precision Count beads (BioLegend) were added to 100 μl of P1 BM sample to allow absolute quantification of hematopoietic cell subsets, and red blood cell lysis was performed.…”
Section: Clinical Flow Cytometry For Bm Analysis/compositionmentioning
confidence: 99%
“…The detailed protocol for whole BM staining has been previously reported (Basso-Ricci et al, 2017). In brief, Precision Count beads (BioLegend) were added to 100 μl of P1 BM sample to allow absolute quantification of hematopoietic cell subsets, and red blood cell lysis was performed.…”
Section: Clinical Flow Cytometry For Bm Analysis/compositionmentioning
confidence: 99%
“…In spite of these observations, studies on the phenotypic composition and transcriptome of these spontaneously mobilized HSPCs were lacking in literature. To assess relative frequencies and absolute counts/µl of blood cellular components, we took advantage of a multiparametric flow-cytometry analysis that allows the dissection of 23 different blood cell types, including HSPCs, myeloid and lymphoid progenitors 27 . Notably, HSC counts/µl were comparable to those observed in the bone marrow of healthy children, supporting the feasibility of exploiting non-mobilized peripheral blood as an easily accessible source for hematopoietic stem cells in these patients.…”
Section: Discussionmentioning
confidence: 99%
“…We further characterized the composition of ARO PB CD34+ cells by deep multiparametric phenotyping, since CD34+ cells are composed by subpopulations with distinct differentiation and survival capability. Blood samples from 5 ARO patients (age range 5.5-8 months, mean 6.8 months) were analyzed using whole blood dissection (WBD) technology, a novel flow cytometry protocol, which allows to evaluate and quantify all major hematopoietic lineages including 10 distinct HSPC subpopulations circulating in the periphery 27 . As reference, we also analyzed peripheral blood samples from 4 age-matched pediatric healthy donors (age range 4-7 months, mean 5.3 months) and bone marrow samples of 7 pediatric healthy donors (age range 3-17 years, mean 10.5 years) ( Figure 1B).…”
Section: High Number Of Circulating Stem/progenitor Cells In Tcirg1-dmentioning
confidence: 99%
“… The lifespan and the counts of peripheral blood cells were obtained from the literature, whereas for MPP counts we applied allometric scaling from rat to human given the specific marker‐dependency of flow cytometry data. Then we derived CMP and MEP counts using the proportions between these populations and MPP, which are experimentally reported for humans ( Supplementary Text ). Mean transit times in humans were calculated from the literature according to the number of non proliferative transit compartments in our model. The drug‐dependent parameters, denoted Slope , were scaled using species‐specific in vitro drug potency experimental results ( Table and Supplementary Text ), which express the drug effect on several proliferative cell types.…”
Section: Methodsmentioning
confidence: 99%
“…QSP Model for HaematotoxicityFornari et al for humans21 (Supplementary Text S9). Mean transit times in humans were calculated from the literature5 according to the number of non proliferative transit compartments in our model.…”
mentioning
confidence: 99%