Superparamagnetic iron oxide labeling and its effects on biological characteristics of neural stem cells
- VernacularTitle:超顺磁性氧化铁标记神经干细胞及对其生物学特性的影响
- Author:
Zhaoyan WANG
;
Yinxiang YANG
;
Zuo LUAN
- Publication Type:Journal Article
- From:
Chinese Journal of Tissue Engineering Research
2007;0(10):-
- CountryChina
- Language:Chinese
-
Abstract:
BACKGROUND: Cell labeling and nuclear magnetic resonance can non-invasively in vivo label the region, existing mode and some bionomics of transplanted neural stem cells (NSCs). OBJECTIVE: To observe the outcome of superparamagnetic iron oxide in vitro labeled human NSCs. DESIGN, TIME AND SETTING: The cytology, in vitro, observation study was performed at the Laboratory of Department of Pediatrics, Navy General Hospital of Chinese PLA from December 2006 to June 2007. MATERIALS: Aborted human embryo was provided by Navy General Hospital of Chinese PLA. Superparamagnetic iron oxide (Lot number 97060601) was produced by Advanced magnetics,inc., USA. METHODS: Monoplast suspension was isolated from human embryo hippocampus using the mechanical method, and in vitro incubated in NSC medium, supplemented with epidermal growth factor and basic fibroblast growth factor. 11.2 g/L superparamagnetic iron oxide was diluted into 28 mg/L using NSC medium, mixed with 1.0 mg/L polylysine (8 ?L), and then made into superparamagnetic iron oxide-polylysine composite labeled medium. NSC spheres with active proliferation were obtained, made into single cell suspension (1?109/L), and then treated with serum-free medium containing superparamagnetic iron oxide. One week later, various cytokines were removed, and 5% fetal bovine serum was used for 24 hours to induce the NSC differentiation. MAIN OUTCOME MEASURES: Prussian blue staining was utilized to determine marking positive rate. Immunofluorescence staining was applied to detect glial fibrillary acidic protein and neurofilament expression. RESULTS: Superparamagnetic iron oxide labeled NSCs were yellow, and the speed of clone formation was not stepped down compared with the non-labeled cells. 20 hours following superparamagnetic iron oxide labeling, blue iron particles in NSCs cytoplasm were found, with the positive rate of 90%. Following induction, superparamagnetic iron oxide labeled NSCs were positive for glial fibrillary acidic protein and neurofilament. CONCLUSION: Superparamagnetic iron oxide can highly effectively label NSCs in vitro, and not affect biological features following labeling. NSCs can normally amplify and orientedly differentiate into neurons and astrocytes.
