Successful use of HTF as a basal fertilization medium during SEcuRe mouse in vitro fertilization.
Assisted reproductive technologies
Cryopreservation
Genetic engineering
In vitro fertilization
Mouse models
Sperm
Journal
BMC research notes
ISSN: 1756-0500
Titre abrégé: BMC Res Notes
Pays: England
ID NLM: 101462768
Informations de publication
Date de publication:
24 Aug 2023
24 Aug 2023
Historique:
received:
09
03
2023
accepted:
08
08
2023
medline:
28
8
2023
pubmed:
25
8
2023
entrez:
24
8
2023
Statut:
epublish
Résumé
The ever-increasing number of genetically engineered mouse models highlights the need for efficient archiving and distribution of these lines. Sperm cryopreservation has become the preferred technique for the majority of these models due to its low requirement of costs, time, and experimental animals. Yet, current in vitro fertilization (IVF) protocols either exhibit decreased fertilization efficiency for the most popular C57BL/6 strain, as recently demonstrated by us, or require costly and difficult-to-prepare media, respectively. As a result, we previously developed SEcuRe, a modified IVF protocol with low costs and high fertilization efficiency. The popular basal fertilization medium, Cook's Here we show that human tubal fluid (HTF), a popular and widely available medium with a known composition, can be used as a basal fertilization medium instead of RVF. Comparison of RVF and HTF during 58 independent SEcuRe IVFs with cryopreserved C57BL/6 sperm revealed equal fertilization and live birth rates. In addition, we demonstrate that HTF has a substantially extended shelf-life by utilizing commercial HTF that was six months past its expiration date, yet did not affect fertilization during IVF or subsequent embryo development. This finding not only increases the economic value of our modified method, but also validates it once more. Our results demonstrate that common, shelf-life extended HTF can be used in SEcuRe IVF in place of now-discontinued RVF medium and ensure the applicability of the method, which we since termed SEcuRe 2.0. Our modified SEcuRe 2.0 strategy will assist researchers to efficiently archive and distribute genetically engineered mouse models in a cost-effective, easily adaptable, and 3R-compliant manner with minimal animal use.
Identifiants
pubmed: 37620881
doi: 10.1186/s13104-023-06452-6
pii: 10.1186/s13104-023-06452-6
pmc: PMC10463834
doi:
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Pagination
184Subventions
Organisme : Deutsche Forschungsgemeinschaft
ID : KFO 329
Informations de copyright
© 2023. BioMed Central Ltd., part of Springer Nature.
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