title: Cytokine Dosage in Fresh and Preserved Human Amniotic Membrane.
abstract: OBJECTIVE
  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.
authors.affiliation: *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.
authors.forename: Mário Genilhu
authors.initials: MG
authors.lastname: Bomfim Pereira
authors.forename: José Alvaro
authors.initials: JA
authors.lastname: Pereira Gomes
authors.forename: Luiz Vicente
authors.initials: LV
authors.lastname: Rizzo
authors.forename: Priscila Cardoso
authors.initials: PC
authors.lastname: Cristovam
authors.forename: Laila Christina
authors.initials: LC
authors.lastname: Silveira
mesh_terms: D000650:Amnion / Q000378:metabolism*; D016207:Cytokines / Q000378:metabolism*; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D005500:Follow-Up Studies; D006801:Humans; D011247:Pregnancy; D011446:Prospective Studies; D013997:Time Factors; D014021:Tissue Preservation / Q000379:methods*
chemical_list: D016207:Cytokines