Effect of accutase dissociation and passage on the apoptosis of human striatum derived neural stem cells.
10.11817/j.issn.1672-7347.2015.05.003
- Author:
Ting LI
1
;
Xiaoli WANG
;
Juan SONG
;
Chen LI
;
Cuiying ZHANG
;
Jie ZHAO
;
Jinsheng WANG
Author Information
1. Department of Physiology, Changzhi Medical College, Changzhi Shanxi 046000, China.
- Publication Type:Journal Article
- MeSH:
Apoptosis;
Caspase 3;
metabolism;
Cells, Cultured;
Collagenases;
chemistry;
Corpus Striatum;
cytology;
Female;
Humans;
In Situ Nick-End Labeling;
Neural Stem Cells;
cytology;
Peptide Hydrolases;
chemistry;
Pregnancy
- From:
Journal of Central South University(Medical Sciences)
2015;40(5):471-478
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVE:To explore the status of apoptosis in human striatum derived neural stem cells (NSCs) aft er accutase dissociation and passage.
METHODS:The NSCs were isolated from fetuses obtained through spontaneous abortion at 13- 18 weeks of pregnancy, which formed neurospheres in vitro. At passages of 3-5, the neurospheres were disassociated into single cell by accutase digestion and then passaged. At 1, 24 and 72 h after passage, the apoptosis of NSCs was measured by several methods, including active caspase-3 or TUNEL staining for fixed cells, Annexin V, Hoechst or PI staining for live cells.
RESULTS:At all of the 3 time points, the staining of TUNEL and active caspase-3 overlapped perfectly. The apoptosis rate of NSCs increased significantly from 20%-25% at 1 h to 75%-80% at 24 h after passage (P<0.01). At 72 h, the apoptosis rate was significantly decreased as compared to that at 24 h time point because of the self-renewal and proliferation of survived NSCs (P<0.01).
CONCLUSION:Many cells in the neurospheres formed by human striatum-derived NSCs underwent apoptosis soon after accutase disassociation. For NSCs cultured in vitro, anti-apoptosis treatments might be a good method to increase the self-renewal and the proliferation of NSCs.
