International Journal of Cell Cloning, Vol 7, 322-329, Copyright © 1989 by AlphaMed Press
Capillary cloning of primary human tumor cells: assay miniaturization for drug efficacy testing
XE Peng, GZ Chen, YF Lu and MJ Murphy Jr
Hipple Cancer Research Center, Dayton, Ohio 45439-2092.
The conventional double-layer agar method of cloning human tumor cells
requires a substantial number of viable tumor cells and 14-21 days of
culture. These prerequisites frequently limit its utility as an assay. In
an attempt to circumvent these limitations and to reduce the amount of drug
that is needed in the assay, we have further developed and miniaturized the
assay in which human tumor cells are cloned in glass microcapillary tubes.
Cultures consisted of 50 microliters containing 15,000 nucleated cells in
975 mm capillary tubes which were incubated for seven days. The results
from 50 consecutive tumor biopsies resulted in cloning efficiencies,
ranging from 0.007% to 1.0% with an overall successful cloning of 88% of
all tumors tested and a good linear growth relationship and chemotherapy
sensitivity. This miniaturized assay offers distinct advantages for drug
efficacy testing including high cloning efficiencies, small tumor sample
and drug requirements, quicker assay turnaround time and a general
conservancy of reagents and incubator space.
