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M.S. Hoonjan, D.A. Johnson, N. Kumar, R.D. Glickman; Amniotic Membrane Effects on Growth Rate of a Melanoma Cell Line . Invest. Ophthalmol. Vis. Sci. 2006;47(13):2721.
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Amniotic membrane transplantation is used in ocular surface reconstruction due to its ability to reduce ocular surface inflammation and to promote epithelialization due to the presence of growth factors. It has been used successfully in ocular surface reconstruction following tumor excisions. This study is an attempt to determine if amniotic membrane transplantation has any effect on the growth rate of a melanoma cell line in vitro.
A specimen of AmnioGraftTM amniotic membrane was obtained from Bio–Tissue, Inc. and irrigated with sterile balanced saline solution (BSS). The specimen was removed from its nitrocellulose filter paper backing with a wet weight of 10.7 mg. The sample was homogenized in a Virtis Virtishear and suspended in 2 mL of cell growth media. Dilutions of the suspended amniotic membrane were added to individual wells in a 24–well plate containing 500 cells per well of the ATCC A2058 human melanoma cell line. A control group with no amniotic membrane, and groups of 5 uL, 10 uL, 15 uL, 20 uL and 25 uL of suspended amniotic membrane were created. Each day, the Pierce BCA protein assay was combined with lysed melanoma cells to determine total protein amounts. The plates were read with a Packard SpectraCount plate reader.
ANOVA using Bonferroni’s method showed a statistically significant growth of the control melanoma cell lines (p=0.009). Similar analysis showed no statistically significant influence on growth rates of the melanoma cells in all of the specified groups from the amniotic membrane over the course of 3 days.
In this limited study, the melanoma cell line did not appear to have any inhibitory or stimulation effect from the introduction of amniotic membrane. The highly transformed melanoma cell line may not have sufficient growth regulatory mechanisms to respond to amniotic tissue growth factors. Further study will be needed to determine if these results are applicable to other cell lines.
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