Please use this identifier to cite or link to this item: http://dx.doi.org/10.18419/opus-1959
Authors: Klünder, Irene
Hülser, Dieter F.
Title: β-galactosidase activity in transfected Ltk- cells is differentially regulated in monolayer and in spheroid cultures
Issue Date: 1993
metadata.ubs.publikation.typ: Zeitschriftenartikel
metadata.ubs.publikation.source: Experimental cell research 207 (1993), S. 155-162
URI: http://nbn-resolving.de/urn:nbn:de:bsz:93-opus-69125
http://elib.uni-stuttgart.de/handle/11682/1976
http://dx.doi.org/10.18419/opus-1959
Abstract: We have investigated whether three-dimensional cultivation of cells to multicell spheroids influences the expression of a transfected gene. Ltk- cells (mouse fibroblasts. thymidine kinase negative) have been transfected with a bacterial lacZ gene which was coupled to a β-actin promoter. The transfected cells synthesize β-galactosidase, a cytoplasmic enzyme which can easily be stained for histology with 5-bromo-4-chloro-3-indoxyl β-D-galactoside and for cytometry with fluorescein di(β-D-galactopyranoside). As we have shown with monolayer cells, β-galactosidase is produced independently of cell density, medium condition, and cell cycle. In multicell spheroids, however, the portion of producing cells was reduced from ~98% to ~2% within a week. This reduction is also independent of cell density, medium condition, and cell cycle. Nonproducing multicell spheroid cells, however, regained their ability to synthesize β-galactosidase within a few days when the cells were recultivated as monolayers. Since the lacZ gene was not lost, its expression might have been regulated by its β-actin promoter. We, therefore, investigated whether the endogenous synthesis of β-actin was similarly regulated. A correlation between the distinct reduction in β-galactosidase-producing cells and filamentous or total actin concentration was not unequivocally observed.
Appears in Collections:04 Fakultät Energie-, Verfahrens- und Biotechnik

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