Evaluating the Effects of Fresh and Cryopreserved Amniotic Membrane on Viability of HeLa and MDA-MB-231 Cancer Cells and Angiogenesis of Rat Aorta Ring

Previous studies have shown that the amniotic membrane and its cells can be an appropriate choice for cancer treatment due to their anticancer properties. This research was designed to evaluate the impact of cryopreservation method on the anti-angiogenic and apoptosis induction properties of amniotic membrane.
In this study, the cancer cells were treated with fresh and cryopreserved amniotic membrane condition medium during 24 hours and the percentage of cancer cells viability was determined using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. To evaluate the changes of angiogenesis, the rat aorta ring assay was examined for both fresh and cryopreserved amniotic membrane within 7 days, and penetration and lack of penetration of fibroblasts-like and capillary-like cells of the aorta were evaluated in both amniotic epithelial and mesenchymal sides of fresh and cryopreserved amniotic membrane, in the presence and absence of epithelial cells.
Findings: Viability of cultured cancer cells treated with condition medium of fresh and cryopreserved amniotic membrane decreased dose-dependently and no significant difference was observed between the fresh and cryopreserved amniotic membrane. The aorta ring assay in fresh and cryopreserved amniotic membrane revealed that the amniotic epithelial stem cells inhibited the penetration of fibroblast-like cells and angiogenesis. Moreover, the penetration of fibroblast-like cells was observed in both epithelial and mesenchymal sides of fresh and cryopreserved amniotic membrane, after the removing epithelial cells.
According to the results of this study, cryopreserved amniotic membrane condition medium reduced the viability of cancer cells, as well as the fresh amniotic membrane condition medium. Moreover, it seems that the maintenance of epithelial cells layer and basement membrane of the amniotic membrane preserves the angiomodulatory properties of amniotic membrane in the cryopreservation method.