A new method for study of normal lens developmentin vitro using pulsatile delivery of PDGF or EGF in HL-1 serum-free medium |
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Authors: | Barbara Brewitt John I Clark |
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Institution: | (1) Department of Biological Structure SM20, University of Washington, 98195 Seattle, Washington;(2) National Eye Institute, National Institutes of Health, Bldg. 6 Rm. 208, 20892 Bethesda, Maryland |
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Abstract: | Summary A new culture method was used to study increases in wet and dry weight and soluble protein during normal development of the
transparent lens. Seven different media with more than ten different additives were tested for their effects on cultured lens
transparency.In vivo, rat lenses increased 53% in soluble protein content between 3 and 5.5 days of age. Only HL-1 serum-free medium containing
15 μg/ml insulin plus 1–2 ng/ml BB platelet-derived growth factor (PDGF), or 5–7 ng/ml epidermal growth factor (EGF) allowed
similar growthin vitro, during the same time period. Normal lens grwoth occurred in culture when fresh medium was delivered to lenses as a pulse
every 4–6 hours. Lenses decreased in dry weight and soluble protein content, and became opaque when the same medium was delivered
continuously. Lenses increased only 26% and 32% in soluble protein content when delivered pulses of HL-1 medium containing
BB PDGF or EGF in the absence of insulin. We suggest that pulsatile delivery of medium containing insulin and PDGF or EGF
stimulates lens growth during developmentin vitro. This pulsatile organ culture system is presented as a new approach for studying the effects of growth factors on cell proliferation,
differentiation, and receptor regulation in a developing tissue.
This work was supported by grants from EY-07031 and EY-04542 from the National Eye Institute and a grant from the Oculon Corporation.
Editor's statement This report documents an in vitro system that may mimic lens development and response to growth regulators
and hormones. The system may be useful for application to other organs and provide a foundation for cell and molecular level
analysis. |
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Keywords: | serum-free media lens culture pulsatile delivery PDGF EGF lens development transparency |
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