Viability and developmental potential of porcine blastocysts preserved for short term in a chemically defined medium at ambient temperature

Abstract

This study developed an efficient method for liquid storage of in vitro‐derived porcine blastocysts at ambient temperature for 24 hr. We evaluated the effects of new chemically defined media (cell wash and preservation solution, Cellstor®‐W [Cell‐W] and cell suspension and preservation solution, Cellstor®‐S [Cell‐S]) for short‐term storage. In the first experiment, in vitro‐derived blastocyst were stored at 25ºC for 24 hr in Cell‐W solution, Cell‐S solution and pig embryo culture (PBM) medium. There were no differences in the rates of survival and development of stored blastocysts between the Cell‐S and Cell‐W solutions, but the total cell number of embryos that survived after storage in Cell‐S solution was significantly higher (p < .05) than that in the Cell‐W solution. In the second experiment, Cell‐S solution was used to store the in vitro‐derived blastocysts at 20°C, 25°C and 30°C. Storage at 20°C resulted in a significant decrease in the rates of survival and development of stored blastocysts compared to storage at 25°C or 30°C. No differences in survival and development rates were observed between storage at 25°C and 30°C, but the damage to the embryo quality after storage and culture was significantly lower at 25°C than at 30°C. In the third experiment, Cell‐S solution was supplemented with β‐mercaptoethanol and curcumin, either alone or in combination, as antioxidant agents. Although the supplementation with curcumin did not improve survival, it significantly increased the development rate of stored blastocysts compared with the control blastocysts stored without antioxidants. In conclusion, when porcine blastocysts were stored at 25°C for 24 hr, a Cell‐S solution may be effective for maintaining the survival and development of in vitro embryos.

Comparison of chemically mediated CRISPR/Cas9 gene editing systems using different nonviral vectors in porcine embryos

“…In vitro fertilization (IVF) was carried out as described previously (Lin et al, 2022). Frozen–thawed boar spermatozoa (Landrace breed) were transferred into 5 mL of porcine fertilization medium (PFM; Research Institute for the Functional Peptides Co., Yamagata, Japan) and washed by centrifugation at 500× g for 5 min.…”

Avoiding aggregation of human bone marrow–derived mesenchymal stem cells stored in cell preservation solutions

“…Dextran, a component of LR-3T-5D, suppresses cell sedimentation and thereby helps maintain a stable concentration of cells in the infusion line (Fujita et al 2020 ). This solution was shown to improve the preservation of porcine embryos (Lin et al 2022a ; Lin et al 2022b ), and the addition of dimethyl sulfoxide or propylene glycol allows solutions to be used for cryopreservation (Fujita et al 2021 ) of, for example, porcine bone marrow–derived MSCs (Nishimura et al 2019 ; Kikuchi et al 2021 ).…”