Noninvasive estimation of cell cycle phase and proliferation rate of human mesenchymal stem cells by phase-shifting laser microscopy.
The simultaneous determination of the cell cycle phase of individual adherent mesenchymal stem cells (MSCs) using a fluorescence microscope after staining with 4',6-diamidine-2'-phenylindole dihydrochloride and bromodeoxyuridine and the laser phase shift by phase-shifting laser microscopy (PLM) revealed that the laser phase shift of cells in the G(2)/M phase was markedly higher than that of cells in the G(0)/G(1) phase. Even in the synchronous cultures to G(0)/G(1) and G(2)/M cell cycle phases, the laser phase shift of the cells in the G(2)/M phase was markedly higher than that of the cells in the G(0)/G(1) phase. The analysis of the cultures of MSCs from different donors with the addition of FGF2 at different concentrations revealed that there was a marked negative correlation between the average phase shift and mean generation time. In conclusion, it is possible to estimate noninvasively the proliferation activity of MSCs population by measuring the phase shift using PLM.
Tokumitsu
Ayako
A
Division of Biotechnology and Macromolecular Chemistry, Graduate School of Engineering, Hokkaido University, N13W8 Kita-ku, Sapporo, 060-8628, Japan.
Wakitani
Shigeyuki
S
Takagi
Mutsumi
M
J Biol Chem. 2002 Jul 26;277(30):27449-67
11956189
Cytotechnology. 2003 Nov;43(1-3):89-96
19003212
Biotechnol Lett. 2008 Jul;30(7):1189-95
18338220
Cell Tissue Res. 1980;209(2):177-86
7190467
J Biomed Opt. 2007 Sep-Oct;12(5):054010
17994898
Appl Opt. 2002 Mar 1;41(7):1308-14
11900008
Biotechnol Bioeng. 1993 Feb 20;41(4):429-39
18609571
Int J Biochem Cell Biol. 2007;39(3):597-605
17113814
Biotechnol Lett. 2009 Jan;31(1):39-42
18850071