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.
Shen
Jun
J
Department of Radiology, The Second Affiliated Hospital, Sun Yat-sen University, 107 Yanjiang Road West, Guangzhou 510120, Guangdong, China. junshenjun@hotmail.com
Cheng
Li-Na
LN
Zhong
Xiao-Mei
XM
Duan
Xiao-Hui
XH
Guo
Ruo-Mi
RM
Hong
Guo-Bing
GB
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