International Journal of Cell Cloning, Vol 7, 360-372, Copyright © 1989 by AlphaMed Press
Use of gel-well culture chambers as a liquid culture system to measure responses of hematopoietic colony-forming cells to growth factors
GN Schwartz
Transplantation Laboratory, American Red Cross, Rockville, Maryland 20855.
This report presents the results of an investigation in which Gel-Well
culture chambers were evaluated for their utility as a liquid culture assay
system to measure the responses of hematopoietic colony-forming cells (CFC)
to recombinant and cell-derived growth factors. Gel-Wells, designed for
anchorage-independent cell growth and diffusion of media components,
permitted the weekly replacement of media and growth factors without
removing cells from the culture chambers. In these studies, changes in
cellularity and CFC content in Gel-Well cultures of human umbilical cord
blood cells induced by recombinant interleukin 3 (rIL-3) were quantified.
After one week in culture without rIL-3, the number of erythroid
burst-forming units (BFU-e) had decreased to 25 +/- 38% of pre-values. In
contrast, addition of rIL-3 induced an increase in the number of BFU-e to
390 +/- 135% of pre-values. By three weeks with rIL-3, the number of
granulocyte-macrophage colony-forming units (CFU-gm) had increased to 292
+/- 58% of pre-values. Also, the presence of a bone marrow stromal cell
layer under the Gel-Well helped to maintain the survival of CFC in liquid
culture. These studies demonstrated that Gel-Well culture chambers provide
a useful liquid culture system for studying the responses of CFC to growth
factors.
