Isolation, culture, identification and labeling of neural stem cells from newborn guinea pigs

Ying WANG; Mingmin DONG; Ying DU

ABSTRACT

BACKGROUND:

Transplantation of neural stem cells into inner ear brings hope for treatment of sensorineural deafness, while guinea pig is the preferred animal in developing animal models of aminoglycosides poisoning-induced deafness. Successful animal models have been established after study for several scores of years. How to isolate neural stem cells and label them from hippocampi of guinea pigs is one of the problems to be solved in transplantation of stem cells into inner ear.

OBJECTIVE:

To isolate and culture neural stem cells from hippocampi of newborn guinea pigs so as to lay basis for transplantation of neural stem cell in the treatment of sensorineural deafness.

DESIGN:

Single sample observation

SETTING:

Staff Room of Microbiology and Immunology, Zhengzhou University MATERIALS This experiment was carried out at the Staff Room of Microbiology and Immunology, Zhengzhou University from March to June 2005. Newbornguinea pigs born within 24 hours of either gender, weighing 50 to 80 g, provided by Experimental Animal Center of Zhengzhou University were used in this experiment.

METHODS:

Neural stem cells from the hippocampal tissue of newborn guinea pigs were isolated and cultured by serum-free medium containing basic fibroblast growth factor (bFGF) and epidermal growth factor (EGF).Expression of nidogen was detected with immunocytochemical staining.Neural stem cells were labeled by DAPI.MAIN OUTCOME

MEASURES:

① In vitro growth of neural stem cells; ② neural stem cells nidogen expression was identified by immunofluorescent staining; ③ labeled stem cells by fluorescence and detected the labeling rate.

RESULTS:

①Clone-like cluster of neural stem cells were obtained from hippocampal tissue ② Neural stem cells could express nidogen. ③Labeling rate of DAPI was 93.4% and no attenuation of fluorescence brightness as observed after 8 generations in culture.

CONCLUSION:

The cells obtained from hippocampal tissues of newborn guinea pigs can be used as donor cells in the treatment of neurosensory deafness by transplanting neural stem cells, for their obvious proliferation ability and high-labeled efficiency with fluorescent dye.