Wang Xiuli, Rosol Michael, Ge Shundi, Peterson Denise, McNamara George, Pollack Harvey, Kohn Donald B, Nelson Marvin D, Crooks Gay M
Division of Research Immunology/BMT, Department of Radiology, and Congresman Dixon Cellular Imaging Core, Childrens Hospital Los Angeles, 4650 Sunset Blvd, Los Angeles, CA 90027,USA.
Blood. 2003 Nov 15;102(10):3478-82. doi: 10.1182/blood-2003-05-1432. Epub 2003 Aug 28.
The standard approach to assess hematopoietic stem cell (HSC) engraftment in experimental bone marrow transplantation models relies on detection of donor hematopoietic cells in host bone marrow following death; this approach provides data from only a single time point after transplantation for each animal. In vivo bioluminescence imaging was therefore explored as a method to gain a dynamic, longitudinal profile of human HSC engraftment in a living xenogeneic model. Luciferase expression using a lentiviral vector allowed detection of distinctly different patterns of engraftment kinetics from human CD34+ and CD34+CD38- populations in the marrow NOD/SCID/beta 2mnull mice. Imaging showed an early peak (day 13) of engraftment from CD34+ cells followed by a rapid decline in signal. Engraftment from the more primitive CD34+CD38- population was relatively delayed but by day 36 increased to significantly higher levels than those from CD34+ cells (P <.05). Signal intensity from CD34+CD38-engrafted mice continued to increase during more than 100 days of analysis. Flow cytometry analysis of bone marrow from mice after death demonstrated that levels of 1% donor cell engraftment could be readily detected by bioluminescence imaging; higher engraftment levels corresponded to higher image signal intensity. In vivo bioluminescence imaging provides a novel method to track the dynamics of engraftment of human HSC and progenitors in vivo.
在实验性骨髓移植模型中，评估造血干细胞（HSC）植入的标准方法依赖于在宿主死亡后检测其骨髓中的供体造血细胞；这种方法仅能为每只动物提供移植后单个时间点的数据。因此，人们探索了体内生物发光成像技术，以在活体异种模型中获取人类HSC植入的动态纵向数据。使用慢病毒载体表达荧光素酶，能够检测到骨髓NOD/SCID/β2mnull小鼠中人类CD34+和CD34+CD38-群体截然不同的植入动力学模式。成像显示，CD34+细胞植入的早期峰值出现在第13天，随后信号迅速下降。更为原始的CD34+CD38-群体的植入相对延迟，但到第36天时，其水平显著高于CD34+细胞（P<.05）。在超过100天的分析过程中，CD34+CD38-植入小鼠的信号强度持续增加。对死亡小鼠骨髓进行的流式细胞术分析表明，生物发光成像能够轻松检测到1%供体细胞植入水平；更高的植入水平对应更高的图像信号强度。体内生物发光成像提供了一种追踪人类HSC及其祖细胞在体内植入动态的新方法。
