Accurate quantification of fourteen normal bone marrow cell subsets in infants to the elderly by flow cytometry

Abstract

Background: Studies of normal bone marrow (BM) cell composition by flow cytometry are scarce. Presently, we aimed to quantify 14 cell subsets from infants to elderly patients.Methods: Cell subsets in BM samples from 180 individuals without morphologically abnormal leukocytes were analyzed using a single combination of eight antibodies: CD3/CD10/CD38/CD19/CD36/CD16/CD34/CD45.Results: By comparison with the Holdrinet score, we first validated the immature granulocyte/neutrophil (IGRA/N) ratio as a readily obtainable criterion of BM sample purity in 145 cases. Then, the 115 highly pure samples were selected (IGRA/N ≥ 1.2) and analyzed according to age group. CD34+ myeloblasts became progressively more infrequent with age: median 1.4% in infancy to 0.5% in the elderly. Neutrophils increased: 10.7% to 22.8%; all other myeloid subsets, IGRA, eosinophils, basophils and monocytes remained stable: respectively 40.3% to 46.7%, 2.0% to 2.8%, 0.2% to 0.3%, and 4.4% to 5.0% throughout life. Erythroblasts were lower in children (8.4% to 10.3%) than in adults (12.5% to 15.1%). For lymphoid cells, hematogones and transitional B-cells decreased: 15.5% to 0.6% and 3.6% to 0.1%, respectively; mature lymphocytes remained stable: B-cells: 1.4% to 2.8%, T-cells: 5.8% to 8.7%, and NK-cells: 0.7% to 1.4%. Plasma cells varied slightly: 0.1% to 0.5%. Differences of about 40% were seen in moderately pure (IGRA/N: 0.5 to 1.2) BM samples.Conclusion: We thus provide the first values for 14 myeloid and lymphoid subsets characterizing BM cell composition in 5 age ranges. They should provide important information when screening patients for hematological disorders or abnormal bone marrow development.

The prognostic significance of hematogones and CD34+ myeloblasts in bone marrow for adult B-cell lymphoblastic leukemia without minimal residual disease

“…However, hematogones were detected in part of the patients, and their percentages in nucleated cells were remarkably variable. While previous work showed that hematogones were more easily found in infants and adolescents 23 , our study focused on adult ALL patients, which removed the age factor that may significantly affect the level of hematogones.…”

Detection and monitoring of minimal residual disease in acute megakaryoblastic leukemia in children

“…В связи с этим необходимо остановиться подробно на особенностях кроветворения детей младшего возраста. Популяционный состав клеток костного мозга новорожденных и детей в возрасте менее 1 года значительно отличается от костного мозга детей более старшего возраста и взрослых [10]. В табл.…”

“…У 3 пациентов определялись различные варианты химерного гена MLL: 46XY?, t(10;11)(p11q23) [14]/46XY [6], t(10:11) MLL/MLLT10 и t(9;11)(p22;q23) MLL-AF9. У 4 пациентов определялся сложный кариотип: 45XY,-7, t(3;3)(q21q26), 48XY,+21,+21, der(17), 53XX,t(1;7)(q21;p15),der(1),+2,+6,+7,+7,der(7)t(1;7),+8,+10, +19 [8]/46,XX [10] и 50,XX,+6,+7,+8, +13, der(11), add(p15), del(17)(q24.). У 1 пациента цитогенетические аномалии не выявлены: 46XY.…”

“…Эти изменения препятствуют адекватной аспирации костного мозга при пункции, полученные пунктаты зачастую имеют недостаточный объем и клеточность, а пропорция в них бластных клеток может быть занижена. Фиброз костного мозга вкупе с особенностями кроветворения детей 1-2-го года жизни обусловливает сравнительно низкий процент опухолевых бластных клеток в костном мозге и трудности диагностики при ОМегЛ [8][9][10].…”

Issue Highlights – November 2019

“…Delineation of normal/activated state and identification of abnormal populations is critical (7). Such constantly growing polychromatic assay protocols require realigned baseline assays different from the more traditional 3-4 color MAb combinations (2,(8)(9)(10)(11).…”