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Updated: Mar 30, 2026
Isolation, Cryopreservation and Culture of Human Amnion Epithelial Cells for Clinical Applications
Published on: December 21, 2014
Cytokine Dosage in Fresh and Preserved Human Amniotic Membrane
Mário Genilhu Bomfim Pereira1, José Alvaro Pereira Gomes, Luiz Vicente Rizzo
1*Department of Ophthalmology, Federal University of São Paulo, São Paulo, Brazil; †Albert Einstein Israelite Hospital, São Paulo, Brazil; and ‡Department of Ophthalmology, Federal University of Minas Gerais.
Purpose:
To evaluate cytokine concentrations in amniotic membrane (AM) preserved in different preservation media, temperatures, and times and to compare them with those in fresh AM.
Methods:
Placentas were harvested from 8 women undergoing cesarean delivery, with each then divided into 17 pieces for the following preservation methods: at 2 different temperatures (-80 and 0°C), in 2 different preservation media (dimethyl sulfoxide and enriched TC199; Ophthalmos), and for different time periods (for 1, 7, 60, and 180 days). Nonpreserved fresh AM was used as a control. An enzyme-linked immunosorbent assay was performed on the supernatant for detection of the following cytokines: epidermal growth factor, basic fibroblast growth factor, hepatocyte growth factor, keratinocyte growth factor, transforming growth factor-beta, and interleukins 4 and 10, and the findings were assessed by post hoc analysis of variance.
Results:
AM preserved at -80°C showed less decrease in the concentration of 4 cytokines. Three cytokines showed less decrease in AM preserved in the TC199 medium, whereas 1 showed less decrease in AM preserved in dimethyl sulfoxide. After storage, 5 cytokine concentrations remained stable for up to 1 day, 3 remained stable for up to 7 days, and all showed significant loss thereafter.
Conclusions:
The AM storage temperature of -80°C was found optimal for maintaining the concentrations of most of the tested cytokines, and enriched TC199 medium was the optimal long-term storage medium for maintaining the concentration of 3 of the cytokines, and with less decrease. When possible, AM should be used within 1 to 7 days after harvesting.
