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


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 1.5T 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 5×10(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.


CONCLUSIONS
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.
Department of Radiology, The Second Affiliated Hospital, Sun Yat-sen University, 107 Yanjiang Road West, Guangzhou 510120, Guangdong, China. junshenjun@hotmail.com
Jun
J
Shen
Li-Na
LN
Cheng
Xiao-Mei
XM
Zhong
Xiao-Hui
XH
Duan
Ruo-Mi
RM
Guo
Guo-Bing
GB
Hong
D000818:Animals; D058948:Cell Tracking / Q000379:methods*; D002478:Cells, Cultured; D003287:Contrast Media; D005456:Fluorescent Dyes; D019786:Gadolinium DTPA; D009474:Neurons / Q000166:cytology*; D009930:Organic Chemicals; D011817:Rabbits; D013194:Staining and Labeling / Q000379:methods; D013234:Stem Cells / Q000166:cytology*
D003287:Contrast Media; D005456:Fluorescent Dyes; D009930:Organic Chemicals; C070080:PKH 26; D019786:Gadolinium DTPA