Molecular and Histological Evaluation of Sheep Ovarian Tissue Subjected to Lyophilization
Cryopreservation is routinely used to preserve cells and tissues; however, long time storage brings many inconveniences including the use of liquid nitrogen. Freeze-drying could enable higher shelf-life stability at ambient temperatures and facilitate transport and storage. Currently, the possibilit...
| Main Authors: | , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
MDPI AG
2021-11-01
|
| Series: | Animals |
| Subjects: | |
| Online Access: | https://www.mdpi.com/2076-2615/11/12/3407 |
| _version_ | 1797506915234742272 |
|---|---|
| author | Daniela Bebbere Amir Arav Stefano Mario Nieddu Giovanni Pietro Burrai Sara Succu Pasquale Patrizio Sergio Ledda |
| author_facet | Daniela Bebbere Amir Arav Stefano Mario Nieddu Giovanni Pietro Burrai Sara Succu Pasquale Patrizio Sergio Ledda |
| author_sort | Daniela Bebbere |
| collection | DOAJ |
| description | Cryopreservation is routinely used to preserve cells and tissues; however, long time storage brings many inconveniences including the use of liquid nitrogen. Freeze-drying could enable higher shelf-life stability at ambient temperatures and facilitate transport and storage. Currently, the possibility to freeze-dry reproductive tissues maintaining vitality and functions is still under optimization. Here, we lyophilized sheep ovarian tissue with a novel device named Darya and a new vitrification and drying protocol and assessed effects on tissue integrity and gene expression. The evaluation was performed immediately after lyophilization (Lio), after rehydration (LR0h) or after two hours of in vitro culture (IVC; LR2h). The tissue survived lyophilization procedures and maintained its general structure, including intact follicles at different stages of development, however morphological and cytoplasmic modifications were noticed. Lyophilization, rehydration and further IVC increasingly affected RNA integrity and caused progressive morphological alterations. Nevertheless, analysis of a panel of eight genes showed tissue survival and reaction to the different procedures by regulation of specific gene expression. Results show that sheep ovarian tissue can tolerate the applied vitrification and drying protocol and constitute a valid basis for further improvements of the procedures, with the ultimate goal of optimizing tissue viability after rehydration. |
| first_indexed | 2024-03-10T04:39:14Z |
| format | Article |
| id | doaj.art-1015560159b847a29cd00f7df8218a2c |
| institution | Directory Open Access Journal |
| issn | 2076-2615 |
| language | English |
| last_indexed | 2024-03-10T04:39:14Z |
| publishDate | 2021-11-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Animals |
| spelling | doaj.art-1015560159b847a29cd00f7df8218a2c2023-11-23T03:26:26ZengMDPI AGAnimals2076-26152021-11-011112340710.3390/ani11123407Molecular and Histological Evaluation of Sheep Ovarian Tissue Subjected to LyophilizationDaniela Bebbere0Amir Arav1Stefano Mario Nieddu2Giovanni Pietro Burrai3Sara Succu4Pasquale Patrizio5Sergio Ledda6Department of Veterinary Medicine, University of Sassari, 07100 Sassari, ItalyFertileSAFE Ltd., 11 Haharash St, Ness Ziona 7403118, IsraelDepartment of Veterinary Medicine, University of Sassari, 07100 Sassari, ItalyDepartment of Veterinary Medicine, University of Sassari, 07100 Sassari, ItalyDepartment of Veterinary Medicine, University of Sassari, 07100 Sassari, ItalyFertileSAFE Ltd., 11 Haharash St, Ness Ziona 7403118, IsraelDepartment of Veterinary Medicine, University of Sassari, 07100 Sassari, ItalyCryopreservation is routinely used to preserve cells and tissues; however, long time storage brings many inconveniences including the use of liquid nitrogen. Freeze-drying could enable higher shelf-life stability at ambient temperatures and facilitate transport and storage. Currently, the possibility to freeze-dry reproductive tissues maintaining vitality and functions is still under optimization. Here, we lyophilized sheep ovarian tissue with a novel device named Darya and a new vitrification and drying protocol and assessed effects on tissue integrity and gene expression. The evaluation was performed immediately after lyophilization (Lio), after rehydration (LR0h) or after two hours of in vitro culture (IVC; LR2h). The tissue survived lyophilization procedures and maintained its general structure, including intact follicles at different stages of development, however morphological and cytoplasmic modifications were noticed. Lyophilization, rehydration and further IVC increasingly affected RNA integrity and caused progressive morphological alterations. Nevertheless, analysis of a panel of eight genes showed tissue survival and reaction to the different procedures by regulation of specific gene expression. Results show that sheep ovarian tissue can tolerate the applied vitrification and drying protocol and constitute a valid basis for further improvements of the procedures, with the ultimate goal of optimizing tissue viability after rehydration.https://www.mdpi.com/2076-2615/11/12/3407lyophilizationovarian tissuevitrificationrehydrationgene expressionhistology |
| spellingShingle | Daniela Bebbere Amir Arav Stefano Mario Nieddu Giovanni Pietro Burrai Sara Succu Pasquale Patrizio Sergio Ledda Molecular and Histological Evaluation of Sheep Ovarian Tissue Subjected to Lyophilization Animals lyophilization ovarian tissue vitrification rehydration gene expression histology |
| title | Molecular and Histological Evaluation of Sheep Ovarian Tissue Subjected to Lyophilization |
| title_full | Molecular and Histological Evaluation of Sheep Ovarian Tissue Subjected to Lyophilization |
| title_fullStr | Molecular and Histological Evaluation of Sheep Ovarian Tissue Subjected to Lyophilization |
| title_full_unstemmed | Molecular and Histological Evaluation of Sheep Ovarian Tissue Subjected to Lyophilization |
| title_short | Molecular and Histological Evaluation of Sheep Ovarian Tissue Subjected to Lyophilization |
| title_sort | molecular and histological evaluation of sheep ovarian tissue subjected to lyophilization |
| topic | lyophilization ovarian tissue vitrification rehydration gene expression histology |
| url | https://www.mdpi.com/2076-2615/11/12/3407 |
| work_keys_str_mv | AT danielabebbere molecularandhistologicalevaluationofsheepovariantissuesubjectedtolyophilization AT amirarav molecularandhistologicalevaluationofsheepovariantissuesubjectedtolyophilization AT stefanomarionieddu molecularandhistologicalevaluationofsheepovariantissuesubjectedtolyophilization AT giovannipietroburrai molecularandhistologicalevaluationofsheepovariantissuesubjectedtolyophilization AT sarasuccu molecularandhistologicalevaluationofsheepovariantissuesubjectedtolyophilization AT pasqualepatrizio molecularandhistologicalevaluationofsheepovariantissuesubjectedtolyophilization AT sergioledda molecularandhistologicalevaluationofsheepovariantissuesubjectedtolyophilization |