In vitro establishment of a neural stem cell aging model and investigation of its biological
characteristics

doi:10.3969/j.issn.2095-4344.2012.49.022

Abstract

Abstract:

BACKGROUND: Stem cell aging is mainly manifested by the decreased proliferation and differentiation ability, shortened telomere length, increased telomerase activity and increased expression of aging-related gene and protein.
OBJECTIVE: To establish a neural stem cell aging model in vitro and to explore the aging-related biological characteristics and mechanisms.
METHODS: The third generation of neural stem cells were isolated and purified from the hippocampus of newborn Sprague-Dawley rats, and the cells were cultured in the medium containing 5% CO2 and neural stem cells under 37 ℃ for 2 hours (control group), then 100 mol/L tert-butyl hydroperoxide was added to the cells for 2 hours to establish the neural stem cell aging model in vitro (model group).
RESULTS AND CONCLUSION: Compared with the control group, after being treated with tert-butyl hydroperoxide for 2 hours, the number and volume of neurospheres forming by proliferated neural stem cells significantly decreased and the number of differentiated cells was significantly decreased in the model group; the absorbance was detected by MTT, the quantity of neurospheres and the numerical density of neurons were significantly decreased by 26%, 48% and 61%, respectively; the percentage of senescence-associated β-galactosidase glucosidase-positive neurospheres was increased by 19 times, and the expression levels of p16INK4a and p21Cip1/Waf1 mRNA were significantly increased by 137% and 68%. These results suggest that tert-butyl hydroperoxide can be used to establish the aging model in vitro. p16INK4 and p21Cip1/Waf1 may play a key role in regulating aging process induced by tert-butyl hydroperoxide through signal transduction pathway of p16/Rb and p19ARF/ p53/ p21cip1.

CLC Number:

R394.2

Peng Bin, Wang Chao-li, Feng Li, Wang Ya-ping. In vitro establishment of a neural stem cell aging model and investigation of its biological
characteristics[J]. Chinese Journal of Tissue Engineering Research, 2012, 16(49): 9241-9246.

Figures/Tables 4

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