A simple flow-cytometry method to evaluate peripheral blood contamination of bone marrow aspirates

Delgado, José A.; Guillén-Grima, Francisco; Moreno, Cristina; Panizo, Carlos; Pérez-Robles, Carmen; Mata, Juan José; Moreno, Laura; Arana, Paula; Chocarro, Silvia; Merino, Juana

doi:10.1016/j.jim.2016.12.006

Cited by 28 publications

(22 citation statements)

References 10 publications

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“…Firstly, we describe progressively lower plasma cell content with each pull of bone marrow aspirate. This is consistent with clinical experience and previous studies showing increased hemodilution in later aspirate pulls [32, 33]. Bone marrow aspirates for research are commonly the final pull, and involve a greater volume, typically 10–20 ml.…”

Section: Discussionsupporting

confidence: 88%

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Baseline identification of clonal V(D)J sequences for DNA-based minimal residual disease detection in multiple myeloma

et al. 2019

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Tracking of clonal immunoglobulin V(D)J rearrangement sequences by next generation sequencing is highly sensitive for minimal residual disease in multiple myeloma. However, previous studies have found variable rates of V(D)J sequence identification at baseline, which could limit tracking. Here, we aimed to define the factors influencing the identification of clonal V(D)J sequences. Bone marrow mononuclear cells from 177 myeloma patients underwent V(D)J sequencing by the LymphoTrack assays (Invivoscribe). As a molecular control for tumor cell content, we sequenced the samples using our in-house myeloma panel myTYPE. V(D)J sequence clonality was identified in 81% of samples overall, as compared with 95% in samples where tumor-derived DNA was detectable by myTYPE. Clonality was detected more frequently in patients with lambda-restricted disease, mainly because of increased detection of kappa gene rearrangements. Finally, we describe how the tumor cell content of bone marrow aspirates decrease gradually in sequential pulls because of hemodilution: From the initial pull used for aspirate smear, to the final pull that is commonly used for research. In conclusion, baseline clonality detection rates of 95% or higher are feasible in multiple myeloma. Optimal performance depends on the use of good quality aspirates and/or subsequent tumor cell enrichment.

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Section: Discussionsupporting

confidence: 88%

“…Evaluation by flow cytometry is commonly performed on second pull aspirates, which carry a greater risk of hemodilution. At the same time, flow cytometry is well known to underestimate both plasma cell content and total white blood cells, due to various technical factors and properties of the cells [32, 33].…”

Section: Resultsmentioning

confidence: 99%

Baseline identification of clonal V(D)J sequences for DNA-based minimal residual disease detection in multiple myeloma

et al. 2019

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“…Besides, cases where samples had been considered as ‘hemodiluted’ in the laboratory report were excluded from the study. Concerning this topic, although some recommendations have been published, there is no universally accepted consensus about its correct evaluation . As our study was retrospective, there were no uniform criteria used for this evaluation among the participating laboratories.…”

Section: Methodsmentioning

confidence: 93%

Normal variation of bone marrow B‐cell precursors according to age – reference ranges for studies in myelodysplastic syndromes in Brazil

Lorand-Metze

Longhini

Oliveira-Duarte

et al. 2017

Cytometry Part B Clinical

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“…Thus, significantly lower total B-cell and nPC numbers were detected in hemodiluted vs. non-hemodiluted BM samples, at the expense of decreased percentages of both stages I and II BCP and both CD19 + and CD19 − nPC, but with similar numbers of TBC/NBC. Altogether, these results indicate that BM hemodilution is associated with significantly decreased numbers of BM-derived B-cells (e.g., BCP) and nPC due to the very low numbers of both cell populations in (steady-state) adult blood [33,34], independently of the time point at which the sample was obtained during the course of disease therapy. To the best of our knowledge, this is the first study in which the impact of hemodilution on the distribution of BM B-cell and PC subsets was investigated in MM and shows that accurate analysis of the distribution of normal/residual B-cells requires the assessment of sample quality.…”

Section: Discussionmentioning

confidence: 99%

B-Cell Regeneration Profile and Minimal Residual Disease Status in Bone Marrow of Treated Multiple Myeloma Patients

Pontes

Flores‐Montero

Sanoja-Flores

et al. 2021

Cancers

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B-cell regeneration during therapy has been considered as a strong prognostic factor in multiple myeloma (MM). However, the effects of therapy and hemodilution in bone marrow (BM) B-cell recovery have not been systematically evaluated during follow-up. MM (n = 177) and adult (≥50y) healthy donor (HD; n = 14) BM samples were studied by next-generation flow (NGF) to simultaneously assess measurable residual disease (MRD) and residual normal B-cell populations. BM hemodilution was detected in 41 out of 177 (23%) patient samples, leading to lower total B-cell, B-cell precursor (BCP) and normal plasma cell (nPC) counts. Among MM BM, decreased percentages (vs. HD) of BCP, transitional/naïve B-cell (TBC/NBC) and nPC populations were observed at diagnosis. BM BCP increased after induction therapy, whereas TBC/NBC counts remained abnormally low. At day+100 postautologous stem cell transplantation, a greater increase in BCP with recovered TBC/NBC cell numbers but persistently low memory B-cell and nPC counts were found. At the end of therapy, complete response (CR) BM samples showed higher CD19− nPC counts vs. non-CR specimens. MRD positivity was associated with higher BCP and nPC percentages. Hemodilution showed a negative impact on BM B-cell distribution. Different BM B-cell regeneration profiles are present in MM at diagnosis and after therapy with no significant association with patient outcome.

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A simple flow-cytometry method to evaluate peripheral blood contamination of bone marrow aspirates

Cited by 28 publications

References 10 publications

Baseline identification of clonal V(D)J sequences for DNA-based minimal residual disease detection in multiple myeloma

Baseline identification of clonal V(D)J sequences for DNA-based minimal residual disease detection in multiple myeloma

Normal variation of bone marrow B‐cell precursors according to age – reference ranges for studies in myelodysplastic syndromes in Brazil

B-Cell Regeneration Profile and Minimal Residual Disease Status in Bone Marrow of Treated Multiple Myeloma Patients

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