[Culture and identification of breast cancer stem cells].
OBJECTIVE
To isolate breast cancer stem cells from breast cancer patients and identify their biological characteristics.


METHODS
Mammospheric cells were purified and enriched from the tumor tissues of breast cancer patients using mammosphere culture. Their expressions of CD44 and CD24 were analyzed by flow cytometry, and ALDH1, ESA and Oct4 expressions were determined by Western Blotting. The primary mammospheric and adherent cells, at the density of 2×10(4), 2×10(5) or 2×10(6), were inoculated into NOD/SCID mice to observe their tumorigenic and metastatic activities.


RESULTS
With mammosphere culture method, 62.36% of the mammospheric cells showed CD44(+)/CD24(-/low) phenotype. The expressions of ALDH1, ESA and Oct4 in the mammospheric cells were significantly higher than those in the adherent culture-derived breast cancer cells (P<0.05). Primary mammospheric cells were at least 100-fold more tumorigenic than the adherent cells; the mammospheric cells were associated with liver or lung metastases, but the adherent cells were not.


CONCLUSIONS
Mammosphere culture can be employed to obtain breast cancer stem cells from the tumor tissues of breast cancer patients.
Department of Breast Surgery, Guangzhou Medical College, Guangzhou, China. wenbo_zheng@163.com
Wen-bo
WB
Zheng
Ling-xiao
LX
Pan
Wei
W
Tang
Jin
J
Gao
Xi-gang
XG
Ye
D000328:Adult; D000080924:Aldehyde Dehydrogenase 1 Family; D000818:Animals; D043303:Aryldialkylphosphatase / Q000378:metabolism; D001943:Breast Neoplasms / Q000473:pathology*; D051927:CD24 Antigen / Q000378:metabolism; D018929:Cell Culture Techniques / Q000379:methods*; D005260:Female; D006801:Humans; D018960:Hyaluronan Receptors / Q000378:metabolism; D007527:Isoenzymes / Q000378:metabolism; D051379:Mice; D016688:Mice, Inbred NOD; D016513:Mice, SCID; D008875:Middle Aged; D014411:Neoplastic Stem Cells / Q000166:cytology* / Q000378:metabolism; D050814:Octamer Transcription Factor-3 / Q000378:metabolism; D061251:Primary Cell Culture; D050697:Retinal Dehydrogenase / Q000378:metabolism
D051927:CD24 Antigen; D018960:Hyaluronan Receptors; D007527:Isoenzymes; D050814:Octamer Transcription Factor-3; C494173:POU5F1 protein, human; D000080924:Aldehyde Dehydrogenase 1 Family; C510223:ALDH1A1 protein, human; C515028:ALDH1A1 protein, mouse; D050697:Retinal Dehydrogenase; D043303:Aryldialkylphosphatase; C468406:PON1 protein, human