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. / Graduation date: 1983
| Identifer | oai:union.ndltd.org:ORGSU/oai:ir.library.oregonstate.edu:1957/27137 |
| Date | 27 April 1983 |
| Creators | Shimakawa, Tomoko |
| Contributors | Oh, Suk Y. |
| Source Sets | Oregon State University |
| Language | en_US |
| Detected Language | English |
| Type | Thesis/Dissertation |
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