Slezak S E, Horan P K
Zynaxis Cell Science, Inc. Malvern, PA 19355.
Blood. 1989 Nov 1;74(6):2172-7.
We report a new technology for in vivo tracking of hematopoietic cells, using fluorescent lipophilic probes. Because the probe is irreversibly bound in the lipids of the cell membrane; substantial numbers of dye molecules can be incorporated per cell and thus substantial signal to noise can be achieved. Although this technology can be used for all hematopoietic cells, these first findings are reported on red blood cells (RBCs) owing to the importance of the membrane to RBC function and integrity. We demonstrated that labeling 10% of the RBCs of a rabbit and reinjecting them into the animal makes possible the tracking of these cells at various times after injection. Furthermore, the labeling appears not to affect in vivo cell lifetime or cellular volume changes in response to hypotonic shock. The single cell fluorescence intensity of the labeled RBCs remains relatively constant for 60 days, and an immune response appears not to be generated against labeled cells. That labeled RBCs have lifetime kinetics in vivo, as shown in other studies, indicates that the membranes are functioning normally and are unaltered by the labeling technology. The technology we present is also applicable to white blood cells, bone marrow, and platelets.
我们报告了一种利用荧光亲脂性探针在体内追踪造血细胞的新技术。由于探针不可逆地结合在细胞膜脂质中，每个细胞可掺入大量染料分子，从而可实现高信噪比。尽管该技术可用于所有造血细胞，但由于膜对红细胞功能和完整性的重要性，这些初步研究结果是关于红细胞（RBC）的。我们证明，标记兔10%的红细胞并将其重新注入动物体内，使得在注射后的不同时间追踪这些细胞成为可能。此外，标记似乎不影响体内细胞寿命或细胞对低渗休克的体积变化。标记红细胞的单细胞荧光强度在60天内保持相对恒定，并且似乎不会针对标记细胞产生免疫反应。如其他研究所示，标记红细胞在体内具有寿命动力学，这表明膜功能正常且未被标记技术改变。我们介绍的这项技术也适用于白细胞、骨髓和血小板。
