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Graefe's Archive for Clinical and Experimental Ophthalmology

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01-06-2018 | Basic Science

Cryopreservation of amniotic membrane with and without glycerol additive

Authors: Malina Wagner, Peter Walter, Sabine Salla, Sandra Johnen, Niklas Plange, Stephan Rütten, Tamme W. Goecke, Matthias Fuest

Published in: Graefe's Archive for Clinical and Experimental Ophthalmology | Issue 6/2018

Abstract

Purpose

Amniotic membrane (AM) is an essential tool in ocular surface reconstruction. In this study, we analyzed the differential effects of glycerol and straight storage at − 80 °C for up to 6 months on the structural, biological, and mechanical properties of amniotic membrane (AM).

Methods

Human placentae of 11 different subjects were analyzed. AMs were stored at − 80 °C, either with a 1:1 mixture of Dulbecco’s modified Eagle medium and glycerol (glycerol) or without any medium or additives (straight). Histological image analysis, tensile strength, cell viability, and basic fibroblast growth factor (bFGF) secretion were evaluated at 0.5, 1, 3, and 6 months.

Results

Histologically, neither glycerol nor straight storage significantly altered the epithelial or stromal structure of the AM. However, the cell number of the stroma was significantly reduced during the freezing process, independently of the storage method (p = 0.05–0.001). Tensile strength and Young’s modulus were not influenced by the storage method, but longer storage periods significantly increased the tensile strength of the AMs (p = 0.028). Cell viability was higher in glycerol rather than straight AM samples for up to 3 months of storage (p = 0.047–0.03). Secretion of bFGF at 3 months of storage was significantly higher in glycerol versus straight frozen AM samples (p = 0.04).

Discussion

Glycerol led to higher cell viability and higher bFGF secretion for up to 3 months of AM storage. However, no significant differences between the two methods were observed at 6 months of storage at − 80 °C.

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Title: Cryopreservation of amniotic membrane with and without glycerol additive
Authors: Malina Wagner
Peter Walter
Sabine Salla
Sandra Johnen
Niklas Plange
Stephan Rütten
Tamme W. Goecke
Matthias Fuest
Publication date: 01-06-2018
Publisher: Springer Berlin Heidelberg
Published in: Graefe's Archive for Clinical and Experimental Ophthalmology / Issue 6/2018
Print ISSN: 0721-832X
Electronic ISSN: 1435-702X
DOI: https://doi.org/10.1007/s00417-018-3973-1

At a glance: The ONWARDS insulin icodec trials

Springer Medicine

Cryopreservation of amniotic membrane with and without glycerol additive

Abstract

Purpose

Methods

Results

Discussion

At a glance: The ONWARDS insulin icodec trials

Springer Medicine

Abstract

Purpose

Methods

Results

Discussion

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