Growth, morphology, function, and morphogenetic properties of rat renal glomerular epithelial cells in vitro: effects of retinyl acetate.
We report here that retinyl acetate (RA) regulates growth, morphology, function and cell organization in rat renal glomerular epithelial cells (SGE1). SGE1 cells are able to grow in a serum-free medium (DHFs medium) which is supplemented with insulin, transferrin, selenium, bovine serum albumin (BSA), linoleic acid and epidermal growth factor (EGF). When 0.1 and 1 micrograms/ml RA were added to the medium, the growth rates in the sparse culture were noticeably increased, compared to those in DHFs alone, whereas more than 10 micrograms/ml RA was cytotoxic to the cells. In the confluent culture, addition of 0.1, 1.0 and 10 micrograms/ml RA prolonged the cell survival. Since 10 micrograms/ml RA is not cytotoxic to the confluent culture, the cytotoxic action of RA seems to be dependent on cell density as well as RA dose. Ultrastructural observation revealed that RA treatment caused an increase of microvilli and alteration of cell shape, from flattened to columnar. Biochemical and immunological studies revealed that RA treatment increased the activity of r-glutamyl transpeptidase (GGT) and an amount of the membrane component with molecular mass (Mr) of 108,000 which is identical to one of nephritogenic antigens, Fx1A. By using fluorescence phalloidin stain, it was found that RA treatment increased content and organization of F-actin fibers. Furthermore, in collagen-embedding culture, RA induced 3-dimensional (3D) growth of SGE1 cells leading to the formation of organoids, cystic spheres with central lumen, in a serum-free condition; the addition of DHFs to collagen gel alone was ineffective for the 3D growth.(ABSTRACT TRUNCATED AT 250 WORDS)
Yamada
M
M
Division of Cell Biology, Shigei Medical Research Institute, Okayama, Japan.
Moritoh
C
C
Kawaguchi
M
M
Okigaki
T
T
D000199:Actins / Q000032:analysis; D000818:Animals; D002455:Cell Division / Q000187:drug effects; D002460:Cell Line; D002470:Cell Survival / Q000187:drug effects; D003094:Collagen; D004224:Diterpenes; D004848:Epithelium / Q000187:drug effects / Q000201:enzymology / Q000648:ultrastructure; D026541:Heymann Nephritis Antigenic Complex; D006651:Histocytochemistry; D007678:Kidney Glomerulus / Q000166:cytology* / Q000187:drug effects / Q000201:enzymology / Q000648:ultrastructure; D008562:Membrane Glycoproteins / Q000032:analysis; D008854:Microscopy, Electron; D008856:Microscopy, Fluorescence; D008871:Microvilli / Q000032:analysis / Q000648:ultrastructure; D009024:Morphogenesis; D051381:Rats; D000084562:Retinyl Esters; D014801:Vitamin A / Q000031:analogs & derivatives* / Q000494:pharmacology; D005723:gamma-Glutamyltransferase / Q000378:metabolism
D000199:Actins; D004224:Diterpenes; D026541:Heymann Nephritis Antigenic Complex; D008562:Membrane Glycoproteins; D000084562:Retinyl Esters; D014801:Vitamin A; C009166:retinol acetate; D003094:Collagen; D005723:gamma-Glutamyltransferase