Analysis of the clonal growth and differentiation dynamics of primitive barcoded human cord blood cells in NSG mice
Analysis of the clonal growth and differentiation dynamics of primitive barcoded human cord blood cells in NSG mice
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DOI：
10.1182/blood-2013-06-508432
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发表时间：
2013-10-31
期刊：
影响因子：
20.3
通讯作者：
Eaves, Connie J.
中科院分区：
文献类型：
--
作者：
Cheung, Alice M. S.;Nguyen, Long V.;Eaves, Connie J.
人类脐带血（CB）提供了一个有吸引力的细胞来源，用于临床移植，因为其丰富的细胞含量具有持续的再生能力，尽管细胞的快速重建能力明显不足。然而，非限制性CB移植的克隆动力学仍然知之甚少。为了开始解决这个问题，我们将CD34(+) CB细胞暴露于一个条形码慢病毒文库中，并使用大规模平行测序来量化随后在2只NOD/SCID-IL2R γ(-/-)小鼠的顺序骨髓抽取中检测到的淋巴细胞和髓细胞的克隆分布，每只小鼠移植了大约10(5)个这些细胞，并在2个继发性受体中再移植6个月。在鉴定的196个克隆中，68个在移植后4周被检测到，通常是淋巴细胞。其余的是在移植后13个月的不同时期后检测到的，但随着时间的推移，稳定性普遍增加，其中包括检测到不同谱系的克隆。然而，在一年多的时间里，也获得了个体细胞能够产生T-、B-和骨髓细胞的明确证据，并且这种潜力也能自我更新。这些发现强调了该模型在体内分析人类造血干细胞控制的注意事项和实用性。
Human cord blood (CB) offers an attractive source of cells for clinical transplants because of its rich content of cells with sustained repopulating ability in spite of an apparent deficiency of cells with rapid reconstituting ability. Nevertheless, the clonal dynamics of nonlimiting CB transplants remain poorly understood. To begin to address this question, we exposed CD34(+) CB cells to a library of barcoded lentiviruses and used massively parallel sequencing to quantify the clonal distributions of lymphoid and myeloid cells subsequently detected in sequential marrow aspirates obtained from 2 primary NOD/SCID-IL2R gamma(-/-) mice, each transplanted with similar to 10(5) of these cells, and for another 6 months in 2 secondary recipients. Of the 196 clones identified, 68 were detected at 4 weeks posttransplant and were often lymphomyeloid. The rest were detected later, after variable periods up to 13 months posttransplant, but with generally increasing stability throughout time, and they included clones in which different lineages were detected. However, definitive evidence of individual cells capable of generating T-, B-, and myeloid cells, for over a year, and self-renewal of this potential was also obtained. These findings highlight the caveats and utility of this model to analyze human hematopoietic stem cell control in vivo.