Graduate Thesis Or Dissertation
 

Effect of zinc supplementation on cell growth and lipoprotein binding in human fibroblast cells

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  • Normal human skin fibroblast cells were used to study the effect of zinc supplementation of the media on cell growth and the competitive binding activity of low density lipoprotein (LDL). Cells were grown in the media containing Dulbecco's Modified-Eagle Medium (DMEM), 5% (v/v) fetal calf serum (FCS), and various levels of zinc. Cell counts and protein determination revealed that there was no stimulatory effect of zinc on the growth of cells, showing a flat growth curve with up to 6 μg/ml zinc supplementation. However, zinc supplementation of greater than 6 μg/ml to the medium appeared to be toxic to the cell and thereby prevented growth. When zinc was removed from the medium using Epoxy-activated Sepharose 6B coupled with iminodiacetate, zinc concentration in the medium was markedly reduced to 0.045 μg/ml from 0.210 μg/ml. The cell growth study using this zinc depleted medium exhibited a growth curve similar to that obtained from the earlier study, suggesting that 0.045 μg/ml of zinc in the control medium was still sufficient to support normal cell growth. For the LDL binding study, cells were grown in the media with various levels of zinc supplementation for 7 days and the competitive binding activity of LDL was determined. When cells were grown in the zinc removed medium with 1.5 μg/ml zinc supplementation, the maximum amounts of ¹²⁵I-LDL bound and internalized in the cells were observed; however, higher levels of zinc supplementation to the growth medium caused decreased ¹²⁵I-LDL binding to the cell receptors. These results suggest that zinc may be involved in the binding of LDL to the receptors.
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