Characterization of a fetal urogenital sinus mesenchymal cell line U4F: secretion of a negative growth regulatory activity.
Mesenchymal cell lines derived from fetal rat urogenital sinus organ cultures have been characterized to establish an in vitro system for addressing growth and differentiation regulatory factors involved in mesenchymal-epithelial interactions during prostate morphogenesis. A continuous cell line was developed and designated U4F. Immunocytochemical analysis showed vimentin intermediate filament content confirming a mesenchymal origin. Previous studies with urogenital sinus organ cultures have reported the expression of a negative growth activity, which is stimulatory to protein synthesis and secretion and alters phenotypic morphology of NBT-II bladder epithelial cells. Subconfluent and confluent U4F monolayers did not produce this growth inhibitory activity. Foci of stacked cells were observed 3 wk postconfluency, which evolved into multicellular spheroids. The negative growth activity was expressed in the conditioned medium coordinate with spheroid formation. Transplanted spheroids continued to express the growth inhibitory activity. Morphologic analysis of spheroids showed a cellular capsule and a core of extracellular matrix. A continuous cell strain (U4F1) with altered phenotypic properties, arose spontaneously from long-term U4F cultures. The U4F1 cell strain did not form spheroids, yet expressed the negative growth activity constitutively in monolayer culture. Analyses of physicochemical, immunological, and biological properties showed the activity is identical in conditioned media from urogenital sinus organ cultures, U4F spheroids, and U4F1 monolayers. Based on the combined properties, this activity cannot be ascribed to previously characterized negative growth factors. The establishment of this mesenchymal cell culture system will aid in the further identification of paracrine-acting growth and differentiation regulatory factors secreted by fetal mesenchyme.
Rowley
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Department of Cell Biology, Baylor College of Medicine, Houston, Texas 77030.
D000818:Animals; D000906:Antibodies; D002460:Cell Line; D003238:Connective Tissue / Q000196:embryology; D003239:Connective Tissue Cells; D003470:Culture Media; D046508:Culture Techniques / Q000379:methods; D004261:DNA Replication; D004847:Epithelial Cells; D004848:Epithelium / Q000187:drug effects; D005333:Fetus; D006131:Growth Inhibitors / Q000096:biosynthesis* / Q000302:isolation & purification / Q000494:pharmacology; D007381:Intermediate Filament Proteins / Q000032:analysis; D009924:Organ Culture Techniques / Q000379:methods; D051381:Rats; D011919:Rats, Inbred Strains; D013936:Thymidine / Q000378:metabolism; D014316:Tritium; D001743:Urinary Bladder; D014566:Urogenital System / Q000166:cytology / Q000196:embryology; D014746:Vimentin / Q000032:analysis
D000906:Antibodies; D003470:Culture Media; D006131:Growth Inhibitors; D007381:Intermediate Filament Proteins; D014746:Vimentin; D014316:Tritium; D013936:Thymidine