Selection of mammalian cells based on their cell-cycle phase using dielectrophoresis

被引:120
作者
Kim, Unyoung [1 ]
Shu, Chih-Wen [4 ]
Dane, Karen Y.
Daugherty, Patrick S. [2 ]
Wang, Jean Y. J. [4 ]
Soh, H. T. [1 ,3 ]
机构
[1] Univ Calif Santa Barbara, Dept Mech Engn, Santa Barbara, CA 93106 USA
[2] Univ Calif Santa Barbara, Dept Chem Engn, Santa Barbara, CA 93106 USA
[3] Univ Calif Santa Barbara, Dept Mat, Santa Barbara, CA 93106 USA
[4] Univ Calif San Diego, Sch Med, Moores Canc Ctr, Dept Med, La Jolla, CA 92093 USA
关键词
cell synchronization; microfluidics; cell sorting; electrokinetics;
D O I
10.1073/pnas.0708760104
中图分类号
O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];
学科分类号
07 ; 0710 ; 09 ;
摘要
An effective, noninvasive means of selecting cells based on their phase within the cell cycle is an important capability for biological research. Current methods of producing synchronous cell populations, however, tend to disrupt the natural physiology of the cell or suffer from low synchronization yields. in this work, we report a microfluidic device that utilizes the dielectrophoresis phenomenon to synchronize cells by exploiting the relationship between the cell's volume and its phase in the cell cycle. The dielectrophoresis activated cell synchronizer (DACSync) device accepts an asynchronous mixture of cells at the inlet, fractionates the cell populations according to the cell-cycle phase (G(1)/S and G(2)/M), and elutes them through different outlets. The device is gentle and efficient; it utilizes electric fields that are 1-2 orders of magnitude below those used in electroporation and enriches asynchronous tumor cells in the G(1) phase to 96% in one round of sorting, in a continuous flow manner at a throughput of 2 x 10(5) cells per hour per microchannel. This work illustrates the feasibility of using laminar flow and electrokinetic forces for the efficient, noninvasive separation of living cells.
引用
收藏
页码:20708 / 20712
页数:5
相关论文
共 32 条
[1]  
Darzynkiewicz Z., 1999, CURRENT PROTOCOLS CE, p8.4.1
[2]   Biological methods for cell-cycle synchronization of mammalian cells [J].
Davis, PK ;
Ho, A ;
Dowdy, SF .
BIOTECHNIQUES, 2001, 30 (06) :1322-+
[3]   Evidence for a size-sensing mechanism in animal cells [J].
Dolznig, H ;
Grebien, F ;
Sauer, T ;
Beug, H ;
Müllner, EW .
NATURE CELL BIOLOGY, 2004, 6 (09) :899-U95
[4]   RELATIONSHIP BETWEEN CELL SIZE AND TIME OF INITIATION OF DNA REPLICATION [J].
DONACHIE, WD .
NATURE, 1968, 219 (5158) :1077-&
[5]   CELL-CYCLE CONTROL BY TIMER AND SIZER IN CHLAMYDOMONAS [J].
DONNAN, L ;
JOHN, PCL .
NATURE, 1983, 304 (5927) :630-633
[6]   CYCLIN - A PROTEIN SPECIFIED BY MATERNAL MESSENGER-RNA IN SEA-URCHIN EGGS THAT IS DESTROYED AT EACH CLEAVAGE DIVISION [J].
EVANS, T ;
ROSENTHAL, ET ;
YOUNGBLOM, J ;
DISTEL, D ;
HUNT, T .
CELL, 1983, 33 (02) :389-396
[7]   Dielectrophoretic sorting of particles and cells in a microsystem [J].
Fiedler, S ;
Shirley, SG ;
Schnelle, T ;
Fuhr, G .
ANALYTICAL CHEMISTRY, 1998, 70 (09) :1909-1915
[8]   Electroporation of cells in microfluidic devices: a review [J].
Fox, M. B. ;
Esveld, D. C. ;
Valero, A. ;
Luttge, R. ;
Mastwijk, H. C. ;
Bartels, P. V. ;
van den Berg, A. ;
Boom, R. M. .
ANALYTICAL AND BIOANALYTICAL CHEMISTRY, 2006, 385 (03) :474-485
[9]   SYNCHRONIZATION OF MAMMALIAN CELLS IN VITRO BY INHIBITION OF DNA SYNTHESIS .2. POPULATION DYNAMICS [J].
GALAVAZI, G ;
BOOTSMA, D .
EXPERIMENTAL CELL RESEARCH, 1966, 41 (02) :438-&
[10]  
Gascoyne PRC, 2002, ELECTROPHORESIS, V23, P1973, DOI 10.1002/1522-2683(200207)23:13<1973::AID-ELPS1973>3.0.CO