In vitro effects of cadmium on two different animal cell models.
The aim of this study was to assess comparatively the effects of cadmium on two different in vitro cell models, a cell line derived from proximal tubule renal cells (LLC-PK1) and haemocytes or blood cells of mussels (Mytilus galloprovincialis). Cells were seeded in 96-well microplates and exposed in vitro to different concentrations of cadmium (CdCl(2)) ranging from 10 to 2000 microM for haemocytes and from 1 to 100 microM for LLC-PK1 cells, added to the culture medium. After 24 h of exposure, different assays were performed on haemocytes: neutral red uptake, phagocytosis of neutral red-stained zymosan, XTT assay, activity of lysosomal acid phosphatase and demonstration of the actin cytoskeleton using TRITC-labeled phalloidin. Cell viability expressed as LC50 was 750 microM when using the neutral red assay and 400 microM with the XTT assay. The phagocytic ability and the activity of acid phosphatase increased significantly in cells treated with Cd in a non dose-dependent manner. Doses of Cd above 100 microM caused disruption of the actin cytoskeleton. In LLC-PK1 cells, cell viability expressed as LC50 was found to be around 40 microM when using the neutral red assay and 50-60 microM with MTT and LDH assays, respectively. These results show that mussel haemocytes are in general more resistant to Cd exposure than LLC-PK1 cells. Furthermore, Cd appears to stimulate phagocytic and lysosomal activities in haemocytes in vitro.
Olabarrieta
I
I
Biologia Zelularra eta Histologia Laborategia, Zoologia eta Animali Zelulen Dinamika Saila, Zientzi Fakultatea, UPV/EHU, 644 PK, E-48080 Bilbo, Basque Country, Spain.
L'Azou
B
B
Yuric
S
S
Cambar
J
J
Cajaraville
M P
MP
D000135:Acid Phosphatase / Q000378:metabolism; D000199:Actins / Q000378:metabolism; D000826:Animal Testing Alternatives; D000818:Animals; D049872:Bivalvia; D002104:Cadmium / Q000633:toxicity*; D002460:Cell Line; D002470:Cell Survival / Q000187:drug effects; D003599:Cytoskeleton / Q000187:drug effects / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D006434:Hemocytes / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D007687:Kidney Tubules, Proximal / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D007770:L-Lactate Dehydrogenase / Q000378:metabolism; D008247:Lysosomes / Q000187:drug effects / Q000201:enzymology; D023421:Models, Animal; D009499:Neutral Red / Q000378:metabolism; D010587:Phagocytosis / Q000187:drug effects; D013778:Tetrazolium Salts / Q000378:metabolism; D015054:Zymosan / Q000378:metabolism
D000199:Actins; D013778:Tetrazolium Salts; D002104:Cadmium; D009499:Neutral Red; D015054:Zymosan; D007770:L-Lactate Dehydrogenase; D000135:Acid Phosphatase