A Study on Optimizing the Cryopreservation Methods for Bama Miniature Pig Semen

Huang

In Vitro Cell.Dev.Biol.-Plant

et al. 2015

Petunia (Petunia × hybrida Vilm.) is an important horticultural crop conserved in the National Genebank of China. Here, a droplet-vitrification cryopreservation protocol was developed for petunia shoot tips. An orthogonal array experiment and additional one-factor experiments were performed to optimize key variables, including the age of in vitro plants, the concentration of sucrose in the preculture solution, the preculture duration, the duration of osmoprotection (2.0 M glycerol and 0.4 M sucrose), the length of exposure to and concentration of plant vitrification solution 2 (30% glycerol, 15% dimethyl sulfoxide, 15% ethylene glycol, and 0.4 M sucrose), and the recovery medium. By using the combined results of the orthogonal and one-factor experiments, the droplet-vitrification procedure for petunia cultivar Niu 2 was formulated efficiently and effectively. The highest regrowth levels were obtained using the following procedure: shoot tips were dissected from in vitro plantlets that were 20 d old, precultured in MS liquid medium with 0.2 M sucrose solution for 2 d, incubated with osmoprotection solution for 30 min at 25°C, cryoprotected with PVS2 for 30 min at 0°C, and rapidly immersed in liquid nitrogen. Cryopreserved shoot tips were then diluted in MS liquid medium with 1.2 M sucrose for 20 min at 25°C and regrown on solidified MS basal medium with half concentration of NH 4 NO 3 , KH 2 PO 4 , KNO 3 , and sucrose. Regrowth levels were as high as 80%. No morphological changes were observed and amplification of 15 simple sequence repeats revealed no genetic alterations after cryopreservation.

Section: Resultsmentioning

confidence: 99%

Identification of a highly successful cryopreservation method (droplet-vitrification) for petunia

Huang

In Vitro Cell.Dev.Biol.-Plant

et al. 2015

Mutation Research/Reviews in Mutation Research

“…Apart from pets, several other domestic species are used, such as horses [399][400][401][402][403][404][405], donkeys [399,406], bulls [407][408][409][410][411], goats [412,413], sheep [414,415] and boars [416,417].…”

Section: Domestic Mammalsmentioning

confidence: 99%

The comet assay in animal models: From bugs to whales – (Part 2 Vertebrates)

Gajski

Žegura

Ladeira

et al. 2019

The comet assay has become one of the methods of choice for the evaluation and measurement of DNA damage. It is sensitive, quick to perform and relatively affordable for the evaluation of DNA damage and repair at the level of individual cells. The comet assay can be applied to virtually any cell type derived from different organs and tissues. Even though the comet assay is predominantly used on human cells, the application of the assay for the evaluation of DNA damage in yeast, plant and animal cells is also quite high, especially in terms of biomonitoring. The present extensive overview on the usage of the comet assay in animal models will cover both terrestrial and water environments. The first part of the review was focused on studies describing the comet assay applied in invertebrates. The second part of the review, (Part 2) will discuss the application of the comet assay in vertebrates covering cyclostomata, fishes, amphibians, reptiles, birds and mammals, in addition to chordates that are regarded as a transitional form towards vertebrates. Besides numerous vertebrate species, the assay is also performed on a range of cells, which includes blood, liver, kidney, brain, gill, bone marrow and sperm cells. These cells are readily used for the evaluation of a wide spectrum of genotoxic agents both in vitro and in vivo. Moreover, the use of vertebrate models and their role in environmental biomonitoring will also be discussed as well as the comparison of the use of the comet assay in vertebrate and human models in line with ethical principles. Although the comet assay in vertebrates is most commonly used in laboratory animals such as mice, rats and lately zebrafish, this paper will only briefly review its use regarding laboratory animal models and rather give special emphasis to the increasing usage of the assay in domestic and wildlife animals as well as in various ecotoxicological studies.

Reprod Domestic Animals

“…Although several authors have studied the effects of different trehalose concentrations, the interaction of trehalose with other cryoprotectants has not been extensively considered (Gutiérrez-Pérez, de Lourdes Juárez-Mosqueda, Carvajal, & Ortega, 2009;Kong et al, 2012;Storey, Noiles, & Thompson, 1998).…”

Section: Introductionmentioning

confidence: 99%

The protease inhibitor antipain has a beneficial synergistic effect with trehalose for ram semen cryopreservation

Akhtarshenas

Shabankareh

Hajarian

et al. 2018

The objective of this study was to examine the different concentrations of antipain and trehalose combination on post-thawed quality of ram semen cryopreserved in tris extender. Ejaculates were collected from four rams using the artificial vagina, pooled at 37°C and diluted with (A Tre : antipain 0 μM and trehalose 0 mM (Control); A Tre ; A Tre ; A Tre ; A Tre ; A Tre ; A Tre ; A Tre and A Tre ). Diluted semen samples were gradually cooled down from 37 to 5°C in a cold cabinet; then, they were loaded into 0.25 ml straws, frozen and stored in liquid nitrogen. Sperm motility (CASA), viability, membrane functionality and abnormality were evaluated after thawing process. Progressive motility in extender supplemented with A Tre , A Tre and A Tre significantly (p < 0.05) higher as compared to the control (A Tre ). A Tre (47.50 ± 0.73) provided the best maintenance of progressive motility in comparison with the control (40.50 ± 0.73). No significant differences were observed between all treated groups in terms of total motility, VAP, VSL, VCL, ALH, BCF, STR and LIN. The percentages of sperm with viable were significantly higher in extenders supplemented with A Tre , A Tre , A Tre and A Tre , compared to control. Addition of A Tre , A Tre and A Tre to extenders improved the percentages of sperm abnormality, compared to the controls. A Tre (67.84 ± 1.51) treatment provided the best maintenance of normal morphology compared to the other treatments. The supplementation with A Tre , A Tre and A Tre improved the percentage of sperm membrane functionality when compared to the control (p < 0.05). Comparing these results with those of control diluents, the effects of supplementation were better except for A Tre group. In conclusion, when combination of antipain (10 μM) and trehalose (30 and 60 mM) was added, they conferred a great cryosurvival capacity with their synergic effects during freeze-thawing process.