Spheres isolated from Colo205 cell line possess cancer stem-like cells under serum-free culture condition / 南方医科大学学报
Journal of Southern Medical University
;
(12): 236-240, 2008.
Article
in Chinese
| WPRIM
| ID: wpr-293408
ABSTRACT
<p><b>OBJECTIVE</b>Isolation and expansion tumor spheres from colorectal cancer cell line Colo205 cultured in serum-free medium(SFM) supplemented with human recombinant EGF and bFGF.</p><p><b>METHODS</b>Colo205 cells were cultivated in SFM,while cells cultivated in serum-supplemented medium(SSM) served as the control. Cells morphology were observed by optical microscope, and expression of intestinal stem cells marker Musashi-1 was detected by immunocytochemical. To induce cell differentiation, tumour spheres were cultivated without EGF and bFGF in the presence of 10% serum. Then we analysed expressions of stem cell surface markers CD133 and CD44 among undifferentiated cell, post-differentiated cells and routine Colo205 cells under serum-supplemented culture condition by flow cytometry. At last we compared cell cycle and spectral karyotype between two groups.</p><p><b>RESULTS</b>In SFM consisting of EGF and bFGF, a minority of Colo205 cells could survive, proliferate and form the suspended tumor spheres. We detected high Musashi-1 expression in these cells. Compared with the SSM group and the post-differentiation SFM group, the expressions of CD133 and CD44 were significantly increased in the undifferentiated SFM group (P<0.05). There was no statistical difference in the expression of CD133 and CD44 between the post-differentiation SFM group and the SSM group (P>0.05). Cell cycle analysis indicated that tumor spheres were of a high proliferation state.We could not find any noticeable difference in the number of chromatosomes between the SFM group and the SSM group.</p><p><b>CONCLUSION</b>Tumor spheres in which enriched cancer stem cells can be generated under serum-free culture condition with EGF and bFGF.</p>
Full text:
Available
Index:
WPRIM (Western Pacific)
Main subject:
Peptides
/
Neoplastic Stem Cells
/
Glycoproteins
/
Antigens, CD
/
RNA-Binding Proteins
/
Culture Media, Serum-Free
/
Spheroids, Cellular
/
Cell Culture Techniques
/
Hyaluronan Receptors
/
Cell Biology
Limits:
Humans
Language:
Chinese
Journal:
Journal of Southern Medical University
Year:
2008
Type:
Article
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