Nanodiamonds for tracking of leukemic cells
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KeywordsLeukemic Cell Trypan Blue K562 Cell Leukemia Cell Line Human Leukemia Cell
Determination of stem cell behavior in vivo is a major challenge in the study of normal and leukemic hematopoiesis. This requires dynamic tracking of cells in vitro in cell culture based assays and in vivo in transplantation models. Most of the existing methods for tracking cells are based on gene transfers that might modulate the engraftment and/or leukemogenicity of transplanted cells . In order to evaluate the use of alternative gene-transfer free methods, we tested in vitro labeling of leukemic cells using nanodiamonds (ND). NDs have become the most promising candidate in recent years due to their excellent biocompatibility, chemical stability, scalability, fluorescent properties and easy functionalization .
Materials and methods
Human leukemia cell lines HL60 and K562, grown in RPMI1640 medium containing 10% fetal bovine serum, were incubated with different concentrations of 6nm nanodiamond - phosphate buffered saline- suspension. Cell viability was assessed using Trypan blue exclusion method at 24h and 72h of incubation. Flow cytometry was performed after 24h and 72 h of incubation to detect the scatter properties of cells. Confocal fluorescence microscopy was performed to detect nanodiamonds after 24 hr incubation.
Our experiments demonstrate for the first time that nanodiamonds can be used successfully in labeling and in vitro tracking of leukemic cell lines using flow cytometry and confocal fluorescence imaging, making them a potential candidate for studying in vivo tracking in xenograft or syngenic mouse models of leukemia.
Authors would like to thank KACST for funding the project (grant number 09BIO-693-03).
This article is published under license to BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.