[A simple method for isolating and culturing single cancer stem cells].
OBJECTIVE
To develop an effective method for isolating and culturing single cancer stem cells.


METHODS
The capillary glass tube was stretched on fire and connected to a sterile plastic tube to prepare the single cell separation apparatus. Single SW480 cell clone spheres in serum-free culture were marked with CD133 and CK7, and the single cancer stem cells were separated and cultivated in 96-well plates or microdrop covered by paraffin.


RESULTS
SW480 cell clone formation rate was about 1.04%, and the cell clone spheres highly expressed CD133 with low CK7 expression. The isolation of the single cancer stem cells showed a success rate of 98.99% using the separation device. The cell division profile was comparable between the cell cultures in microdrop and 96-well plates in the initial 2 cell divisions (P>0.05), whereas prolonged cell division occurred afterwards in the microdrop culture as compared to 96-well plate culture. The cell population expansion of the single cancer stem cells was similar between microdrop culture (11.5%, 22/192) and 96-well plate culture (9.2%, 17/184) (P>0.05).


CONCLUSIONS
Single SW480 cells can develop into cancer stem cell spheres. Microdrop culture is convenient and stable, and can be the primary choice for single cancer stem cell culture.
Department of Pathology, Southern Medical University, Guangzhou, China. dengyj@fimmu.com
Yongjian
Y
Deng
Qiang
Q
Jiang
Lina
L
Yu
Hongxia
H
Wang
Chunting
C
Hu
Chao
C
Liu
Yanqing
Y
Ding
D018929:Cell Culture Techniques / Q000379:methods*; D045744:Cell Line, Tumor; D002469:Cell Separation / Q000379:methods*; D006801:Humans; D014411:Neoplastic Stem Cells / Q000166:cytology*