The Effect of Choline Salt Addition to Trehalose Solution for Long-Term Storage of Dried and Viable Nuclei from Fully Grown Oocytes
Although drying techniques are exciting alternatives to cryopreservation, it remains challenging to maintain tightly controlled temperatures and humidity levels during storage of dried products. The objective of this study was to determine if the addition of choline acetate to trehalose solution cou...
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MDPI AG
2023-08-01
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| Series: | Bioengineering |
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| Online Access: | https://www.mdpi.com/2306-5354/10/9/1000 |
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| author | Joseph A. Orozco Cabral Pei-Chih Lee Shangping Wang Yizhou Wang Yong Zhang Pierre Comizzoli Gloria D. Elliott |
| author_facet | Joseph A. Orozco Cabral Pei-Chih Lee Shangping Wang Yizhou Wang Yong Zhang Pierre Comizzoli Gloria D. Elliott |
| author_sort | Joseph A. Orozco Cabral |
| collection | DOAJ |
| description | Although drying techniques are exciting alternatives to cryopreservation, it remains challenging to maintain tightly controlled temperatures and humidity levels during storage of dried products. The objective of this study was to determine if the addition of choline acetate to trehalose solution could enable a wider range of storage conditions for preservation of nuclei from fully grown oocytes, by allowing temporary humidity excursions (>44% relative humidity) that may lead to crystallization of trehalose and loss of DNA integrity. Using domestic cat germinal vesicle oocytes as a model, we characterized the recovery as well as the integrity of samples after microwave-assisted dehydration. Exposure to choline acetate alone did not impair the germinal vesicle’s DNA integrity and only had a negative impact on the chromatin configuration. Choline acetate addition enabled us to reach lower moisture contents after 25 min of microwave-assisted drying. Sample recovery after rehydration was also better in the presence of choline acetate. The integrity of the germinal vesicle’s DNA was not affected, while the chromatin configuration was impaired by the presence of choline acetate during dehydration. Importantly, choline acetate addition helped to maintain an amorphous state (absence of detrimental crystallization) during excursion from ideal humidity conditions. |
| first_indexed | 2024-03-10T23:03:02Z |
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| id | doaj.art-eeacd95999374c4a90f52d66d56a8f42 |
| institution | Directory Open Access Journal |
| issn | 2306-5354 |
| language | English |
| last_indexed | 2024-03-10T23:03:02Z |
| publishDate | 2023-08-01 |
| publisher | MDPI AG |
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| series | Bioengineering |
| spelling | doaj.art-eeacd95999374c4a90f52d66d56a8f422023-11-19T09:36:17ZengMDPI AGBioengineering2306-53542023-08-01109100010.3390/bioengineering10091000The Effect of Choline Salt Addition to Trehalose Solution for Long-Term Storage of Dried and Viable Nuclei from Fully Grown OocytesJoseph A. Orozco Cabral0Pei-Chih Lee1Shangping Wang2Yizhou Wang3Yong Zhang4Pierre Comizzoli5Gloria D. Elliott6Department of Mechanical Engineering and Engineering Sciences, University of North Carolina at Charlotte, Charlotte, NC 28223, USASmithsonian’s National Zoo and Conservation Biology Institute, Washington, DC 20008, USADepartment of Bioengineering, Clemson University, Clemson, SC 29634, USANanoscale Science Graduate Program, University of North Carolina at Charlotte at Charlotte, Charlotte, NC 28223, USAElectrical and Computer Engineering Department, University of North Carolina at Charlotte, Charlotte, NC 28223, USASmithsonian’s National Zoo and Conservation Biology Institute, Washington, DC 20008, USADepartment of Mechanical Engineering and Engineering Sciences, University of North Carolina at Charlotte, Charlotte, NC 28223, USAAlthough drying techniques are exciting alternatives to cryopreservation, it remains challenging to maintain tightly controlled temperatures and humidity levels during storage of dried products. The objective of this study was to determine if the addition of choline acetate to trehalose solution could enable a wider range of storage conditions for preservation of nuclei from fully grown oocytes, by allowing temporary humidity excursions (>44% relative humidity) that may lead to crystallization of trehalose and loss of DNA integrity. Using domestic cat germinal vesicle oocytes as a model, we characterized the recovery as well as the integrity of samples after microwave-assisted dehydration. Exposure to choline acetate alone did not impair the germinal vesicle’s DNA integrity and only had a negative impact on the chromatin configuration. Choline acetate addition enabled us to reach lower moisture contents after 25 min of microwave-assisted drying. Sample recovery after rehydration was also better in the presence of choline acetate. The integrity of the germinal vesicle’s DNA was not affected, while the chromatin configuration was impaired by the presence of choline acetate during dehydration. Importantly, choline acetate addition helped to maintain an amorphous state (absence of detrimental crystallization) during excursion from ideal humidity conditions.https://www.mdpi.com/2306-5354/10/9/1000microwave-assisted dehydrationoocyte nucleustrehalosecholine acetaterelative humidity |
| spellingShingle | Joseph A. Orozco Cabral Pei-Chih Lee Shangping Wang Yizhou Wang Yong Zhang Pierre Comizzoli Gloria D. Elliott The Effect of Choline Salt Addition to Trehalose Solution for Long-Term Storage of Dried and Viable Nuclei from Fully Grown Oocytes Bioengineering microwave-assisted dehydration oocyte nucleus trehalose choline acetate relative humidity |
| title | The Effect of Choline Salt Addition to Trehalose Solution for Long-Term Storage of Dried and Viable Nuclei from Fully Grown Oocytes |
| title_full | The Effect of Choline Salt Addition to Trehalose Solution for Long-Term Storage of Dried and Viable Nuclei from Fully Grown Oocytes |
| title_fullStr | The Effect of Choline Salt Addition to Trehalose Solution for Long-Term Storage of Dried and Viable Nuclei from Fully Grown Oocytes |
| title_full_unstemmed | The Effect of Choline Salt Addition to Trehalose Solution for Long-Term Storage of Dried and Viable Nuclei from Fully Grown Oocytes |
| title_short | The Effect of Choline Salt Addition to Trehalose Solution for Long-Term Storage of Dried and Viable Nuclei from Fully Grown Oocytes |
| title_sort | effect of choline salt addition to trehalose solution for long term storage of dried and viable nuclei from fully grown oocytes |
| topic | microwave-assisted dehydration oocyte nucleus trehalose choline acetate relative humidity |
| url | https://www.mdpi.com/2306-5354/10/9/1000 |
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