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Cryobiology
Academic Press
Cryobiology

Academic Press

0011-2240

Cryobiology/Journal CryobiologySCIISTP
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    Towards a method for cryopreservation of mosquito vectors of human pathogens

    Gallichotte, Emily N.Dobos, Karen M.Ebel, Gregory D.Hagedorn, Mary...
    10页
    查看更多>>摘要：Mosquito-borne diseases are responsible for millions of human deaths every year, posing a massive burden on global public health. Mosquitoes transmit a variety of bacteria, parasites and viruses. Mosquito control efforts such as insecticide spraying can reduce mosquito populations, but they must be sustained in order to have long term impacts, can result in the evolution of insecticide resistance, are costly, and can have adverse human and environmental effects. Technological advances have allowed genetic manipulation of mosquitoes, including generation of those that are still susceptible to insecticides, which has greatly increased the number of mosquito strains and lines available to the scientific research community. This generates an associated challenge, because rearing and maintaining unique mosquito lines requires time, money and facilities, and long-term maintenance can lead to adaptation to specific laboratory conditions, resulting in mosquito lines that are distinct from their wild-type counterparts. Additionally, continuous rearing of transgenic lines can lead to loss of genetic markers, genes and/or phenotypes. Cryopreservation of valuable mosquito lines could help circumvent these limitations and allow researchers to reduce the cost of rearing multiple lines simultaneously, maintain low passage number transgenic mosquitoes, and bank lines not currently being used. Additionally, mosquito cryopreservation could allow researchers to access the same mosquito lines, limiting the impact of unique laboratory or field conditions. Successful cryopreservation of mosquitoes would expand the field of mosquito research and could ultimately lead to advances that would reduce the burden of mosquito-borne diseases, possibly through rear-and-release strategies to overcome mosquito insecticide resistance. Cryopreservation techniques have been developed for some insect groups, including but not limited to fruit flies, silkworms and other moth species, and honeybees. Recent advances within the cryopreservation field, along with success with other insects suggest that cryopreservation of mosquitoes may be a feasible method for preserving valuable scientific and public health resources. In this review, we will provide an overview of basic mosquito biology, the current state of and advances within insect cryopreservation, and a proposed approach toward cryopreservation of Anopheles stephensi mosquitoes.
      原文链接:
    • NSTL
    • Elsevier

    The effects of broccoli and caraway extracts on serum oxidative markers, testicular structure and function, and sperm quality before and after sperm cryopreservation

    Raeeszadeh, MahdiehKhademi, NadiaAkbari, Abolfazl
    9页
    查看更多>>摘要：Despite studies on the effects of medicinal plants on reproductive performance, the effect of extracts broccoli and caraway on serum and testicular oxidative biomarkers, testicular structure and function and sperm quality before and after cryopreservation has not been studied. Sixty-three male mice were divided into nine controlled and treated groups as follow: control, broccoli (200 mg/kg), broccoli (300 mg/kg), caraway (200 mg/kg), caraway (300 mg/kg), broccoli-caraway (200 mg/kg), broccoli (300 mg/kg)-caraway (200 mg/kg), broccoli (200 mg/ kg)-caraway (300 mg/kg), broccoli-caraway (300 mg/kg). After 42 days of treatment, the animals were sacrificed and blood sample and testicular tissue were collected for biochemical and histological measurements. Sperm quality was also measured before and after cryopreservation. The results showed that the diameter and number of spermatogonium, primary spermatocytes, spermatids and sperm count were significantly increased by broccoli (300 mg/kg), while level of them were significantly decreased by caraway (300 mg/kg) compared to other groups (p < 0.01). Sperm viability and motility after thawing significantly improved by broccoli (300 mg/ kg) compared to control. Testosterone levels significantly increased by broccoli (300 mg/kg) compared to control and caraway (300 mg/kg). The serum and testicular SOD and CAT activity significantly increased by broccoli (300 mg/kg) compared to other groups (p < 0.05). MDA and DNA fragmentation levels significantly increased by caraway (200 and 300 mg/kg) compared to others (p < 0.01). It can be concluded that broccoli extract in a dose dependent manner than caraway extract could improve serum and testes oxidative biomarkers, testicular structure and function, and sperm quality before and after cryopreservation.
      原文链接:
    • NSTL
    • Elsevier

    Investigating solution effects injury of human T lymphocytes and its prevention during interrupted slow cooling

    Liu, WeiHuang, ZhiyongLiu, BaolinHe, Xiaowen...
    8页
    查看更多>>摘要：Cooling rate is a critical parameter affecting the success of cell cryopreservation. Fast cooling can result in intracellular ice formation (IIF), while slow cooling can bring solution effects injury, both are detrimental to the cells. Whilst most of the studies have investigated how IIF affects cells, solution effects injury has received little attention. Here, we studied the solution effects injury of human T lymphocytes by cryomicroscopy and tested the osmoprotective ability of some frequently used cryoprotective agents (CPAs) such as dimethyl sulfoxide (DMSO), glycerol, trehalose, urea and L-proline. We further investigated the relationship between cell volume, latent heat and solution effects cell injury. We found that solution effects injury during interrupted slow cooling was caused by high concentration of the extracellular solution rather than eutectic formation and solutes precipitation. DMSO, glycerol and trehalose can protect cells from solution effects injury, while L-proline and urea cannot under the same condition. The cell volume and latent heat are not crucial for causing solution effects injury in cells. This work confirms that high osmotic pressure, rather than eutectic formation, leads to cell injury. It also suggests that cell volume and latent heat may not be a key factor for explaining solution effects injury and its prevention in the cryopreservation of human T lymphocytes.
      原文链接:
    • NSTL
    • Elsevier

    Ice recrystallization inhibition activity varies with ice-binding protein type and does not correlate with thermal hysteresis

    Graham, Laurie A.Eves, RobertAgrawal, PrashantOleschuk, Richard D....
    12页
    查看更多>>摘要：Ice-binding proteins (IBPs) inhibit the growth of ice through surface adsorption. In some freeze-resistant fishes and insects, circulating IBPs serve as antifreeze proteins to stop ice growth by lowering the freezing point. Plants are less able to avoid freezing and some use IBPs to minimize the damage caused in the frozen state by ice recrystallization, which is the growth of large ice grains at the expense of small ones. Here we have accurately and reproducibly measured the ice recrystallization inhibition (IRI) activity of over a dozen naturally occurring IBPs from fishes, insects, plants, and microorganisms using a modified ?splat? method on serial dilutions of IBPs whose concentrations were determined by amino acid analysis. The endpoint of IRI, which was scored as the lowest protein concentration at which no recrystallization was observed, varied for the different IBPs over two orders of magnitude from 1000 nM to 5 nM. Moreover, there was no apparent correlation between their IRI levels and reported antifreeze activities. IBPs from insects and fishes had similar IRI activity, even though the insect IBPs are typically 10x more active in freezing point depression. Plant IBPs had weak antifreeze activity but were more effective at IRI. Bacterial IBPs involved in ice adhesion showed both strong freezing point depression and IRI. Two trends did emerge, including that basal plane binding IBPs correlated with stronger IRI activity and larger IBPs had higher IRI activity.
      原文链接:
    • NSTL
    • Elsevier

    Cryopreservation of Spix's yellow-toothed cavy epididymal sperm using Tris- and coconut water-based extenders supplemented with egg yolk or Aloe vera

    Moreira, Samara Sandy Jeronimoda Silva, Andreia MariaSouza, Ana Liza PazPraxedes, Erica Camila Gurgel...
    6页
    查看更多>>摘要：Addressing the establishment of biobanks for the conservation of wild hystricomorph rodents? germplasm, we verified the effects of different extenders and distinct concentrations of non-permeant cryoprotectants on the sperm parameters of Spix's yellow-toothed cavies. Nine testis-epididymis complexes were used for sperm collection by retrograde washing using Tris or a powdered coconut water extender (ACP (R)-116c). Spermatozoa were diluted and frozen with the same extenders supplemented with egg yolk or Aloe vera at a 10% or 20% concentration. After recovery and cryopreservation, all samples were evaluated for sperm kinetic parameters, morphology, membrane integrity, osmotic response, and sperm-binding capability using an egg yolk perivitelline membrane assay. After recovery, no differences were observed between Tris and ACP (R)-116c that provided 515.4 x 10(6) sperm/mL and 561.6 x 10(6) sperm/mL, presenting >65% motile sperm, respectively. After cryo-preservation, most effective preservation of sperm kinetic parameters (68.1 x 5.9% motile sperm) and membrane integrity (48.2 x 7.4%) was provided by Tris extender supplemented with 10% egg yolk. However, both ex-tenders supplemented with any concentration of egg yolk or Aloe vera presented similar preservation of osmotic response and sperm-binding ability after cryopreservation. In summary, we suggest the use of a Tris extender supplemented of 10% egg yolk for cryopreservation of Spix's yellow-toothed cavy epidydimal sperm.
      原文链接:
    • NSTL
    • Elsevier

    Antioxidant defense response during hibernation and arousal in Chinese soft-shelled turtle Pelodiscus sinensis juveniles

    Tang, Zhong-huaChen, Bo-jianNiu, Cui-juan
    9页
    查看更多>>摘要：Antioxidant defense is essential for animals to cope with homeostasis disruption during hibernation. The present study aimed to investigate the antioxidant defense response of juvenile soft-shelled turtle Pelodiscus sinensis during hibernation and following arousal. Turtle brain, liver, and kidney samples were collected at pre hibernation (17 degrees C mud temperature; MT), during hibernation (5.8 degrees C MT) and after arousal (20.1 degrees C MT) in the field. Transcript levels of NF-E2-related factor 2 (Nrf2) decreased significantly during hibernation and recovered after arousal in all tissues. Cerebral and nephric copper-zinc superoxide dismutase (Cu/Zn SOD), catalase (CAT), glutathione peroxidase 3 (GPx3) and nephric GPx4 mRNA showed similar changing patterns as Nrf2. Cerebral Mn SOD, GPx1 and nephric GPx1 up-regulated after arousal. Hepatic Cu/Zn SOD, GPx1 and GPx3 mRNA kept stable, except hepatic GPx4 increased during hibernation. Hepatic Mn SOD and CAT increased after arousal. In the GSH system, mRNA levels of glutathione synthetases (GSs) kept stable during hibernation and up regulated after arousal in most tissues except nephric GS2 mRNA remained unchanged. Gene expressions of glutathione reductase (GR) exhibited a tissue specific changing pattern, while those of glutathione-S-Transferase (GST) shared a similar pattern among tissues: remained stable or down-regulated during hibernation then recovered in arousal. In contrast to these diverse responses in gene expressions, most of the antioxidant enzyme activities maintained high and stable. Overall, no preparation for oxidative stress (POS) strategy was found in enzymatic antioxidant system in P. sinensis juveniles during hibernation, the Chinese soft-shelled turtles were able to stay safe from potential oxidative stress during hibernation by maintaining high level activities/concentrations of the antioxidant enzymes/antioxidants.
      原文链接:
    • NSTL
    • Elsevier

    Alteration of the lipid phase transition during mouse embryos freezing after in vitro culture with linoleic acid

    Igonina, T. N.Okotrub, K. A.Brusentsev, E. YuChuyko, E. A....
    9页
    查看更多>>摘要：Lipids significantly affect embryo cryopreservation in some mammalian species depending on the cell lipidome quantity and composition. One of the ways to study the relationship between lipid content and cryotolerance of cells is to study the effect of lipidome modification on laboratory mice. The objective of this research was to study how in vitro culture of mouse embryos with linoleic acid (LA) will affect lipid phase transition (LPT) during cooling and subsequent embryo development after cryopreservation. Embryos obtained in vivo at the 2-cell stage were cultured with 200 mu M LA for 46 h up to the morula/blastocyst stage. Thereafter, one portion of embryos was slowly frozen to reveal the effect of LA on survival after cryopreservation, another portion was used to characterize the lipid composition and to determine the temperature of the LPT onset. Confocal fluorescence microscopy of Nile Red stained embryos showed a significant increase in lipid content of the LA treated group compared to the controls. Raman measurements showed that the onset of LPT in LA treated embryos is lower than in untreated ones: -5 degrees C vs +2 degrees C. However, these changes in the LPT onset did not affect the survival rates of embryos after cryopreservation. In summary, in vitro culture with LA changes the biophysical characteristics of embryos' lipidome and is realized in lower LPT onset, but this does not affect embryo survival after cryopreservation.
      原文链接:
    • NSTL
    • Elsevier

    Differential proteome between ejaculate and epididymal sperm represents a key factor for sperm freezability in wild small ruminants

    Martinez-Fresneda, LuciaSylvester, MarcShakeri, FarhadBunes, Andreas...
    14页
    查看更多>>摘要：Epididymal sperm shows higher cryoresistance than ejaculated sperm. Although the sperm proteome seems to affect cell cryoresistance, studies aiming at identifying proteins involved in sperm freezing-tolerance are scarce. The aims of this study were to investigate differences of sperm freezability and proteome between epididymal and ejaculated sperm in three mountain ungulates: Iberian ibex, Mouflon and Chamois. Sperm samples were cryopreserved in straws by slow freezing. Tandem mass tag-labeled peptides from sperm samples were analyzed by high performance liquid chromatography coupled to a mass spectrometer in three technical replicates. The statistical analysis was done using the moderated t-test of the R package limma. Differences of freezability between both types of sperm were associated with differences of the proteome. Overall, epididymal sperm showed higher freezability than ejaculated sperm. Between 1490 and 1883 proteins were quantified in each species and type of sperm sample. Cross species comparisons revealed a total of 76 proteins that were more abundant in epididymal than in ejaculated sperm in the three species of study whereas 3 proteins were more abundant in ejaculated than epididymal sperm in the three species of study (adjusted P 0.5). Many of the proteins that were associated with higher cryoresistance are involved in stress response and redox homeostasis. In conclusion, marked changes of sperm proteome were detected between epididymal and ejaculated sperm. This work contributes to update the sperm proteome of small ruminants and to identify candidate markers of sperm freezability.
      原文链接:
    • NSTL
    • Elsevier

    The effects of cryopreservation and cold storage on sperm subpopulation structure of common carp (Cyprinus carpio L.)

    Marinovic, ZoranScekic, IlijaLujic, JelenaUrbanyi, Bela...
    7页
    查看更多>>摘要：The objectives of this study were to identify the presence of different spermatozoa subpopulations (SPs) according to their kinematic characteristics in the sperm of common carp and to test the effects of cryopreservation and prolonged (6-day) storage at room temperature (RT; 23 degrees C) and 4 degrees C on spermatozoa motility and subsequently on SP dynamics. Two-step clustering analyses identified three motile SPs based on their kinematic properties: SP1 contained spermatozoa with low velocity and low/moderate STR/LIN values (slow non-linear SP); SP2 was comprised of spermatozoa with high velocities and high STR/LIN values (fast linear SP); SP3 was characterized with high VCL, and moderate LIN/STR (fast non-linear SP); and an additional SP0 was added comprising immotile spermatozoa. Total motility, progressive motility and VCL decreased after cryopreservation to approximately 50% of their value in fresh sperm, while the frequency of SPs characterized by high values of motility parameters declined in favor of those with low motility values and SP0. Motility values of fresh and cryopreserved spermatozoa which were washed with fresh extender after thawing decreased significantly after 24 h of storage at RT and after 72 h of storage at 4 degrees C, while cryopreserved sperm which remained in the original cryomedium faced a steep decline in motility after only 2 h of storage. As subpopulation frequencies followed this dynamic, this indicates that cryopreserved sperm should be washed with fresh extender in order to obtain favorable sperm kinematic properties after freezing.
      原文链接:
    • NSTL
    • Elsevier

    Aseptic capillary vitrification of human spermatozoa: Cryoprotectant-free vs. cryoprotectant-included technologies

    Wang, MengyingTodorov, PlamenIsachenko, EvgeniaRahimi, Gohar...
    8页
    查看更多>>摘要：The protocol of aseptic cryoprotectant-free vitrification on human spermatozoa is well documented. However, data about the effect of permeable cryoprotectants at this procedure is limited. Presented study aimed to test the aseptic capillary vitrification technologies using permeable cryoprotectant-included or cryopmtectant-free media. Thirty-two normal samples were included and analyzed after vitrification in three different media and thawing. Three treatment groups were formed: Group 1, basic medium; Group 2, basic medium with 0.25 M sucrose; Group 3, basic medium with glycerol. Before plunging into liquid nitrogen, capillaries were filled by 10 til of spermatozoa suspension and isolated from liquid nitrogen by location in hermetically closed 0.25 ml straws. Progressive motility, plasma membrane integrity, total motility/viability after 24, 48 and 72 h in vitro culture, apoptosis and mitochondrial membrane potential (Delta Psi m) were determined after thawing at 42 degrees C. Progressive motility of spermatozoa in groups 1, 2, 3 was 24.9 +/- 1.7%, 34.5 +/- 2.8% and 34.0 +/- 1.4%, respectively (P-1-2,P-3 0.1). In conclusion, supplementation of medium for aseptic capillary technology for cryoprotectant-free vitrification of human spermatozoa by permeable cryoprotectant does not improve the quality of spermatozoa after warming.
      原文链接:
    • NSTL
    • Elsevier