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Cryopreservation in a Standard Freezer: −28 °C as Alternative Storage Temperature for Amniotic Membrane Transplantation

Human amniotic membrane (hAM) is usually stored at −80 °C. However, in many regions, cryopreservation at −80 °C is not feasible, making hAM unavailable. Therefore, the possibility of cryopreservation at −28 °C (household freezer) was investigated. hAMs (n = 8) were stored at −80 °C or −28 °C for a m...

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Autores principales: Witt, Joana, Grumm, Luis, Salla, Sabine, Geerling, Gerd, Menzel-Severing, Johannes
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8877302/
https://www.ncbi.nlm.nih.gov/pubmed/35207382
http://dx.doi.org/10.3390/jcm11041109
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author Witt, Joana
Grumm, Luis
Salla, Sabine
Geerling, Gerd
Menzel-Severing, Johannes
author_facet Witt, Joana
Grumm, Luis
Salla, Sabine
Geerling, Gerd
Menzel-Severing, Johannes
author_sort Witt, Joana
collection PubMed
description Human amniotic membrane (hAM) is usually stored at −80 °C. However, in many regions, cryopreservation at −80 °C is not feasible, making hAM unavailable. Therefore, the possibility of cryopreservation at −28 °C (household freezer) was investigated. hAMs (n = 8) were stored at −80 °C or −28 °C for a mean time of 8.2 months. hAM thickness, epithelial integrity and basement membrane were assessed histologically. The collagen content, concentration of hepatocyte growth factor (HGF) and basic fibroblast growth factor (bFGF) were determined. Elastic modulus and tensile strength were measured. The mean thickness of hAM stored at −28 °C was 33.1 ± 21.6 µm (range 9.7–74.9); thickness at −80 °C was 30.8 ± 14.7 µm (range 13.1–50.7; p = 0.72). Mean collagen content, epithelial cell number and integrity score showed no significant difference between samples stored at −28 °C or −80 °C. Basement membrane proteins were well preserved in both groups. Mean tensile strength and elastic modulus were not significantly different. Concentration of bFGF at −28 °C was 1063.2 ± 680.3 pg/g (range 369.2–2534.2), and 1312.1 ± 778.2 pg/g (range 496.2–2442.7) at −80 °C (p = 0.11). HGF was 5322.0 ± 2729.3 pg/g (range 603.3–9149.8) at −28 °C, and 11338.5 ± 6121.8 pg/g (range 4143.5 to 19806.7) at −80 °C (p = 0.02). No microbiological contamination was detected in any sample. The cryopreservation of hAM at −28 °C has no overt disadvantages compared to −80 °C; the essential characteristics of hAM are preserved. This temperature could be used in an alternative storage method whenever storage at −80 °C is unavailable.
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spelling pubmed-88773022022-02-26 Cryopreservation in a Standard Freezer: −28 °C as Alternative Storage Temperature for Amniotic Membrane Transplantation Witt, Joana Grumm, Luis Salla, Sabine Geerling, Gerd Menzel-Severing, Johannes J Clin Med Article Human amniotic membrane (hAM) is usually stored at −80 °C. However, in many regions, cryopreservation at −80 °C is not feasible, making hAM unavailable. Therefore, the possibility of cryopreservation at −28 °C (household freezer) was investigated. hAMs (n = 8) were stored at −80 °C or −28 °C for a mean time of 8.2 months. hAM thickness, epithelial integrity and basement membrane were assessed histologically. The collagen content, concentration of hepatocyte growth factor (HGF) and basic fibroblast growth factor (bFGF) were determined. Elastic modulus and tensile strength were measured. The mean thickness of hAM stored at −28 °C was 33.1 ± 21.6 µm (range 9.7–74.9); thickness at −80 °C was 30.8 ± 14.7 µm (range 13.1–50.7; p = 0.72). Mean collagen content, epithelial cell number and integrity score showed no significant difference between samples stored at −28 °C or −80 °C. Basement membrane proteins were well preserved in both groups. Mean tensile strength and elastic modulus were not significantly different. Concentration of bFGF at −28 °C was 1063.2 ± 680.3 pg/g (range 369.2–2534.2), and 1312.1 ± 778.2 pg/g (range 496.2–2442.7) at −80 °C (p = 0.11). HGF was 5322.0 ± 2729.3 pg/g (range 603.3–9149.8) at −28 °C, and 11338.5 ± 6121.8 pg/g (range 4143.5 to 19806.7) at −80 °C (p = 0.02). No microbiological contamination was detected in any sample. The cryopreservation of hAM at −28 °C has no overt disadvantages compared to −80 °C; the essential characteristics of hAM are preserved. This temperature could be used in an alternative storage method whenever storage at −80 °C is unavailable. MDPI 2022-02-19 /pmc/articles/PMC8877302/ /pubmed/35207382 http://dx.doi.org/10.3390/jcm11041109 Text en © 2022 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Witt, Joana
Grumm, Luis
Salla, Sabine
Geerling, Gerd
Menzel-Severing, Johannes
Cryopreservation in a Standard Freezer: −28 °C as Alternative Storage Temperature for Amniotic Membrane Transplantation
title Cryopreservation in a Standard Freezer: −28 °C as Alternative Storage Temperature for Amniotic Membrane Transplantation
title_full Cryopreservation in a Standard Freezer: −28 °C as Alternative Storage Temperature for Amniotic Membrane Transplantation
title_fullStr Cryopreservation in a Standard Freezer: −28 °C as Alternative Storage Temperature for Amniotic Membrane Transplantation
title_full_unstemmed Cryopreservation in a Standard Freezer: −28 °C as Alternative Storage Temperature for Amniotic Membrane Transplantation
title_short Cryopreservation in a Standard Freezer: −28 °C as Alternative Storage Temperature for Amniotic Membrane Transplantation
title_sort cryopreservation in a standard freezer: −28 °c as alternative storage temperature for amniotic membrane transplantation
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8877302/
https://www.ncbi.nlm.nih.gov/pubmed/35207382
http://dx.doi.org/10.3390/jcm11041109
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