Tissue cryobanking for conservation programs: effect of tissue type and storage time after death

Abstract

In this study, we investigated the temporal post-mortem limits, within which there will be guarantees of obtaining living cells from several tissues of sheep and cattle and the effect of vitrification on the ability of cells from tissue stored at different times. Muscle tissue and auricular cartilage were stored at 4°C for 5, 48, 72, 96 and 216 h post-mortem (hpm). Tissue samples were sorted into two groups: one group was in vitro cultured immediately after storage and the other was vitrified after storage and then in vitro cultured. In cattle and sheep, no differences in subconfluence rates were observed between the two experimental groups. At the same time, no significant differences were observed in the number of days required in culture to reach confluence between non-vitrified and vitrified groups when tissues were stored at 4°C for different times. In sheep, while the population doubling times (PDT) were similar in cartilage cells from vitrified and non-vitrified tissues and stored at 4°C for 5 and 216 hpm, PDT of muscle cells were longer in 216 hpm stored groups than in 5 hpm stored groups. In bovine, although the PDT of muscle cells were similar for 5 and 216 hpm and both vitrified and non-vitrified tissues and the PDT were longer in cartilage cells from vitrified than from non-vitrified tissues. In conclusion, although storage times and vitrification have different effects on tissues from cattle and sheep, this study showed that living cells could be obtained from all groups. Therefore, cartilage and muscle tissues can be stored at 4°C for 216 hpm and used for cyrobanking.

The initial steps toward the formation of somatic tissue banks and cell cultures derived from captive Antillean manatee (Trichechus manatus manatus) skin biopsies

“…In a study by Caamaño et al (2008), vitrification showed lower efficiency compared to SF and noncryopreserved tissues derived from brown bears ( Ursus arctos ), suggesting that high concentrations of cryoprotectants affect the quality of vitrified tissues. Additionally, other factors may influence the efficiency of the SSV, such as the use of a pre‐equilibration solution/step, tissue type (Caputcu et al, 2013), tissue thickness (Carvalho et al, 2013), warming and reanimation conditions (Lima et al, 2018).…”

Conservation of somatic tissue derived from collared peccaries (Pecari tajacu Linnaeus, 1758) using direct or solid-surface vitrification techniques

“…Thus, in regions of difficult access, a greater number of individuals can be collected, without restricting the age and gender, thus allowing an unlimited number of samples and the extrapolation of these cell characteristics for the entire population (León-Quinto et al 2009). Therefore, somatic tissue samples can contribute to preservation programs (Caputcu et al 2013;Silvestre et al 2004). These skin cryobanks permit the maintenance of a maximal representation of biodiversity population, being an important tool in wildlife preservation programs (León-Quinto et al 2014).…”

Use of somatic cell banks in the conservation of wild felids

“…In general, slow freezing uses low concentrations of cryoprotectants and is done through gradual temperature reduction in a controlled manner. Although not performed in wild felids, vitrification has also been a prominent method in tissue cryopreservation and has been used in somatic tissues of some mammals (Borges et al, ; Caputcu, Akkoc, Cetinkaya, & Arat, ). This technique is performed through rapid temperature reduction and with high concentrations of cryoprotectants.…”