Broadening cell selection criteria with micropallet arrays of adherent cells.
A host of technologies exists for the separation of living, nonadherent cells, with separation decisions typically based on fluorescence or immunolabeling of cells. Methods to separate adherent cells as well as to broaden the range of possible sorting criteria would be of high value and complementary to existing strategies. Cells were cultured on arrays of releasable pallets. The arrays were screened and individual cell(s)/pallets were released and collected. Conventional fluorescence and immunolabeling of cells were compatible with the pallet arrays, as were separations based on gene expression. By varying the size of the pallet and the number of cells cultured on the array, single cells or clonal colonies of cells were isolated from a heterogeneous population. Since cells remained adherent throughout the isolation process, separations based on morphologic characteristics, for example cell shape, were feasible. Repeated measurements of each cell in an array were performed permitting the selection of cells based on their temporal behavior, e.g. growth rate. The pallet array system provides the flexibility to select and collect adherent cells based on phenotypic and temporal criteria and other characteristics not accessible by alternative methods.
Wang
Yuli
Y
Department of Physiology and Biophysics, University of California, Irvine, CA 92697, USA.
Young
Grace
G
Aoto
Phillip C
PC
Pai
Jeng-Hao
JH
Bachman
Mark
M
Li
G P
GP
Sims
Christopher E
CE
Allbritton
Nancy L
NL
D000818:Animals; D002448:Cell Adhesion / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects; D002469:Cell Separation; D048430:Cell Shape / Q000187:drug effects; D002999:Clone Cells; D004129:Dimethylpolysiloxanes / Q000494:pharmacology; D005453:Fluorescence; D049452:Green Fluorescent Proteins / Q000378:metabolism; D006367:HeLa Cells; D006801:Humans; D051381:Rats; D013997:Time Factors; D046888:Tissue Array Analysis / Q000379:methods*
D004129:Dimethylpolysiloxanes; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins