4.5 Article

Efficient in vitro labeling rabbit neural stem cell with paramagnetic Gd-DTPA and fluorescent substance

Journal

EUROPEAN JOURNAL OF RADIOLOGY
Volume 75, Issue 3, Pages 397-405

Publisher

ELSEVIER IRELAND LTD
DOI: 10.1016/j.ejrad.2009.04.040

Keywords

Neural stem cells; Magnetic resonance imaging; Transfection; Cell labeling

Funding

  1. National Natural Science Foundation of China [30400115, 30870691]
  2. Natural Science Foundation of Guangdong Province [04300241]

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Objectives: The aim of this study is to label rabbit neural stem cells (NSCs) by using standard contrast agents (Gd-DTPA) in combination with PKH26 and in vitro track them with MR imaging. Materials and methods: NSCs from prenatal brains of rabbits were cultured and propagated. Intracellular uptake of Gd-DTPA was achieved by using a non-liposomal lipid transfection reagent (Effectene) as the transfection agent. After labeling with Gd-DTPA, cells were incubated with cellular membrane fluorescent dye PKH26. The labeling effectiveness and the longevity of Gd-DTPA maintenance were measured on a 1.5 T MR scanner. The influence of labeling on the cellular biological behaviors was assessed by cellular viability, proliferation and differentiation assessment. Results: The labeling efficiency of Gd-DTPA was up to 90%. The signal intensity on T1-weighted imaging and T1 values of labeled cells were significantly higher than those of unlabeled cells (P < 0.05). The minimal number of detectable cells for T1-weighted imaging was 5 x 10(3). Cellular uptake of Gd-DTPA was maintained until 15 days after initially labeling. There was no significant difference in the cellular viability and proliferation between the labeled and unlabeled NSCs (P > 0.05). Normal glial and neuronal differentiation remained in labeled NSCs like unlabeled NSCs. Conclusion: Highly efficient labeling NSCs with Gd-DTPA could be achieved by using Effectene. This method of labeling NSCs allows for tracking cells with MR imaging, and without alterations of cellular biological behaviors. (C) 2009 Elsevier Ireland Ltd. All rights reserved.

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