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Indian Journal of Biochemistry and Biophysics (IJBB) >
IJBB Vol.47 [2010] >
IJBB Vol.47(6) [December 2010] >
| Title: | Isolation of stem-like cells from human MG-63 osteosarcoma cells using limiting dilution in combination with vincristine selection |
| Authors: | Lou, Nan
Wang, Yan
Sun, Dahui
Zhao, Jianwu
Wang, Yang
Gao, Zhongli |
| Keywords: | MG-63 cells
Holoclone
Tumor stem-like cells
Vincristine |
| Issue Date: | Dec-2010 |
| Publisher: | NISCAIR-CSIR, India |
| Abstract: | To isolate stem-like cells from the human MG-63
osteosarcoma cell line, different subpopulations of MG-63 cells were cloned by
limiting dilution and passaged to obtain different sublines. The subline with
highest clonogenicity was identified using a proliferation assay, cell-cycle
analysis, and soft-agar colony-forming assay. The sublines were further
selected in serum-free medium containing 20 ng/ml vincristine to identify
cells that could form suspended sarcospheres. Identified cells were then
characterized based on morphology, cell surface markers, adipogenic and
osteogenic differentiation, and tumorigenicity in nude mice. A total of 19
holoclones that could be stably passaged were obtained. Sublines A1, A3, and D1
were markedly different from other sublines and the parental cell line. Subline
D1 not only had a higher colony-forming efficiency and formed larger colonies,
but also possessed a shorter latency of tumorigenesis in vivo. After subline D1 was cultured in suspension in medium
containing vincristine, a highly enriched subpopulation of cells that could
form sarcospheres and be stably passaged were obtained. These cells, designated
as MG-63-M expressed multiple markers of multipotent or embryonic stem cells
and possessed the capacity for self-renewal, multilineage differentiation, and
significant multi-drug resistance. Thus, our results suggest that a
subpopulation of stem-like cells can be isolated from human MG-63 osteosarcoma
cell line. |
| Page(s): | 340-347 |
| CC License: |  CC Attribution-Noncommercial-No Derivative Works 2.5 India |
| ISSN: | 0975-0959 (Online); 0301-1208 (Print) |
| Source: | IJBB Vol.47(6) [December 2010]
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