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Human bone marrow assessment by single-cell RNA sequencing mass cytometry and flow cytometry

机译:通过单细胞RNA测序大规模流式细胞仪和流式细胞仪评估人类骨髓

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摘要

New techniques for single-cell analysis have led to insights into hematopoiesis and the immune system, but the ability of these techniques to cross-validate and reproducibly identify the biological variation in diverse human samples is currently unproven. We therefore performed a comprehensive assessment of human bone marrow cells using both single-cell RNA sequencing and multiparameter flow cytometry from 20 healthy adult human donors across a broad age range. These data characterize variation between healthy donors as well as age-associated changes in cell population frequencies. Direct comparison of techniques revealed discrepancy in the quantification of T lymphocyte and natural killer cell populations. Orthogonal validation of immunophenotyping using mass cytometry demonstrated a strong correlation with flow cytometry. Technical replicates using single-cell RNA sequencing matched robustly, while biological replicates showed variation. Given the increasing use of single-cell technologies in translational research, this resource serves as an important reference data set and highlights opportunities for further refinement.
机译:单细胞分析的新技术已使人们对造血和免疫系统有了更深入的了解,但是这些技术交叉验证和可再现地鉴定各种人类样品中生物学变异的能力目前尚未得到证实。因此,我们使用单细胞RNA测序和多参数流式细胞术对来自20个健康成年人的广泛年龄范围的人骨髓细胞进行了全面评估。这些数据表征了健康供体之间的变异以及细胞群频率的年龄相关变化。技术的直接比较揭示了T淋巴细胞和自然杀伤细胞群体的定量差异。使用大规模细胞计数法对免疫表型进行正交验证显示出与流式细胞术密切相关。使用单细胞RNA测序的技术复制品具有良好的匹配性,而生物学复制品则表现出差异。鉴于单细胞技术在翻译研究中的使用越来越多,该资源可作为重要的参考数据集,并突出了进一步完善的机会。

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