TY  - JOUR
TI  - Investigating Neural Stem Cell and Glioma Stem Cell Self-renewal Potential Using Extreme Limiting Dilution Analysis (ELDA)
AU  - Nguyen, Hong PT
AU  - Daniel, Paul M
AU  - Filiz, Gulay  
AU  - Mantamadiotis, Theo 
VL  - 8
IS  - 17
PY  - 2018
DA  - 2018/09/05
SP  - e2991
C1  - Bio-protocol 2018;8:e2991
DO  - 10.21769/BioProtoc.2991
UR  - https://doi.org/10.21769/BioProtoc.2991
AB  - Glioma stem cells (GSC) grown as neurospheres exhibit similar characteristics to neural stem cells (NSC) grown as neurospheres, including the ability to self-renew and differentiate. GSCs are thought to play a role in cancer initiation and progression. Self-renewal potential of GSCs is thought to reflect many characteristics associated with malignancy, including tumor recurrence following cytotoxic therapy due to their proliferative dormancy and capacity to allow for the development of resistant tumor cell sub-clones due to mutations acquired during their differentiation. Here, we demonstrate that using extreme limiting dilution analysis (ELDA), subtle differences in the frequency of sphere-forming potential between PI3K-mutant oncogenic NSCs and non-oncogenic NSCs can be measured, in vitro. We further show how ELDA can be used on cells, before and after forced differentiation to amplify inherent differences in sphere-forming potential between mutant and control NSCs. Ultimately, ELDA exploits a difference in the ability of a single or a few seeded stem cells to self-renew, divide and form neurospheres. Importantly, the assay also allows a comparison between genetically distinct cells or between the same cells under different conditions, where the impact of target-specific drugs or other novel cancer stem cell therapies can be tested.
KW  - Stem cells
KW  - Glioma
KW  - Glioblastoma
KW  - Neural stem cells
KW  - Neurospheres
KW  - Self-renewal
KW  - ELDA
JF  - Bio-protocol
SN  - 2331-8325
PB  - Bio-protocol LLC.
BIO101  - False