Browse
Submit Data
Databases
API
Help

Dataset Information

13 Views

0 Connections

0 Citations

0 Reanalyses

0 Downloads

Omics score: 0

Cryopreservation and Laser Nanowarming of Zebrafish Embryos Followed by Hatching and Spawning.

ABSTRACT: This study shows for the first time the ability to rewarm cryopreserved zebrafish embryos that grow into adult fish capable of breeding normally. The protocol employs a single injection of cryoprotective agents (CPAs) and gold nanorods (GNRs) into the yolk and immersion in a precooling bath to dehydrate the perivitelline space. Then embryos are encapsulated within CPA and GNR droplets, plunged into liquid nitrogen, cryogenically stabilized, and rewarmed by a laser pulse. Postlaser nanowarming, embryos (n = 282) exhibit intact structure by 1 h (40%), continued development after 3 h (22%), movement after 24 h (11%), hatching after 48 h (9%), and swimming after Day 5 (3%). Finally, from fish that survives till Day 5, two larvae are grown to adulthood and spawned, yielding survival comparable to an unfrozen control. Future efforts will focus on improving the survival to adulthood and developing methods to cryopreserve large numbers of embryos for research, aquaculture, and biodiversity preservation.

SUBMITTER: Khosla K

PROVIDER: S-EPMC8627598 | biostudies-literature |

REPOSITORIES: biostudies-literature

ACCESS DATA

Json Xml

Similar Datasets

Characterization of Laser Gold Nanowarming: A Platform for Millimeter-Scale Cryopreservation.

Project description:Preventing ice formation during cryopreservation by vitrification has led to the successful storage and banking of numerous cellular- and tissue-based biomaterials. In their breakthrough work, Peter Mazur's group achieved over 90% survival by using a laser warming technique for 100 ?m mice oocytes that were cooled in 0.1 ?L droplets with 2.3 M CPA and extracellularly loaded India ink (laser absorber). Laser warming can provide rapid and uniform warming rates to "outrun" damaging ice crystal growth. Here we generalize Mazur's technique for microliter-sized droplets using laser nanowarming to rewarm millimeter-scale biomaterials when loaded extracellularly and/or intracellularly with biocompatible 1064 nm resonant gold nanoparticles. First, we show that droplets containing low-concentration cryoprotectants (such as 2 M propylene glycol ± 1 M trehalose) can be rapidly cooled at rates up to 90?000 °C/min by plunging into liquid nitrogen to achieve either a visually transparent state (i.e., vitrified) or a cloudy with ice (i.e., nonvitrified) state. Both modeling and experiments were then used to characterize the laser nanowarming process for different laser energy (2-6 J), pulse length (1-20 ms), droplet volume (0.2-1.8 ?L), cryoprotectant (2-3 M), and gold concentration (0.77 × 1017-4.8 × 1017 nps/m3) values to assess physical and biological success. Physical success was achieved by finding conditions that minimize cloudiness and white spots within the droplets during cooling and warming as signs of damaging ice formation and ice crystallization, respectively. Biological success was achieved using human dermal fibroblasts to find conditions that achieve ?90% cell viability normalized to controls postwarming. Thus, physical and biological success can be achieved using this platform cryopreservation approach of rapid cooling and laser gold nanowarming in millimeter-scale systems.

| S-EPMC6536355 | biostudies-literature

Perfusion, cryopreservation, and nanowarming of whole hearts using colloidally stable magnetic cryopreservation agent solutions.

Project description:Nanowarming of cryopreserved organs perfused with magnetic cryopreservation agents (mCPAs) could increase donor organ utilization by extending preservation time and avoiding damage caused by slow and nonuniform rewarming. Here, we report formulation of an mCPA containing superparamagnetic iron oxide nanoparticles (SPIONs) that are stable against aggregation in the cryopreservation agent VS55 before and after vitrification and nanowarming and that achieve high-temperature rise rates of up to 321°C/min under an alternating magnetic field. These SPIONs and mCPAs have low cytotoxicity against primary cardiomyocytes. We demonstrate successful perfusion of whole rat hearts with the mCPA and removal using Custodiol HTK solution, even after vitrification, cryostorage in liquid nitrogen for 1 week, and nanowarming under an alternating magnetic field. Quantification of SPIONs in the hearts using magnetic particle imaging demonstrates that the formulated mCPAs are suitable for perfusion, vitrification, and nanowarming of whole organs with minimal residual iron in tissues.

| S-EPMC7793590 | biostudies-literature

Microencapsulation and nanowarming enables vitrification cryopreservation of mouse preantral follicles.

Project description:Preantral follicles are often used as models for cryopreservation and in vitro culture due to their easy availability. As a promising approach for mammalian fertility preservation, vitrification of preantral follicles requires high concentrations of highly toxic penetrating cryoprotective agents (up to 6 M). Here, we accomplish low-concentration-penetrating cryoprotective agent (1.5 M) vitrification of mouse preantral follicles encapsulated in hydrogel by nanowarming. We find that compared with conventional water bath warming, the viability of preantral follicles is increased by 33%. Moreover, the cavity formation rate of preantral follicles after in vitro culture is comparable to the control group without vitrification. Furthermore, the percentage of MII oocytes developed from the vitrified follicles, and the birth rate of offspring following in vitro fertilization and embryo transfer are also similar to the control group. Our results provide a step towards nontoxic vitrification by utilizing the synergistic cryoprotection effect of microencapsulation and nanowarming.

| S-EPMC9755531 | biostudies-literature

Nanowarming and ice-free cryopreservation of large sized, intact porcine articular cartilage.

Project description:Successful organ or tissue long-term preservation would revolutionize biomedicine. Cartilage cryopreservation enables prolonged shelf life of articular cartilage, posing the prospect to broaden the implementation of promising osteochondral allograft (OCA) transplantation for cartilage repair. However, cryopreserved large sized cartilage cannot be successfully warmed with the conventional convection warming approach due to its limited warming rate, blocking its clinical potential. Here, we develope a nanowarming and ice-free cryopreservation method for large sized, intact articular cartilage preservation. Our method achieves a heating rate of 76.8 °C min-1, over one order of magnitude higher than convection warming (4.8 °C min-1). Using systematic cell and tissue level tests, we demonstrate the superior performance of our method in preserving large cartilage. A depth-dependent preservation manner is also observed and recapitulated through magnetic resonance imaging and computational modeling. Finally, we show that the delivery of nanoparticles to the OCA bone side could be a feasible direction for further optimization of our method. This study pioneers the application of nanowarming and ice-free cryopreservation for large articular cartilage and provides valuable insights for future technique development, paving the way for clinical applications of cryopreserved cartilage.

| S-EPMC9958003 | biostudies-literature

Matrix Metalloproteinase 13 Activity is Required for Normal and Hypoxia-Induced Precocious Hatching in Zebrafish Embryos.

Project description:Hypoxia induces precocious hatching in zebrafish, but we do not have a clear understanding of the molecular mechanisms regulating the activation of the hatching enzyme or how these mechanisms trigger precocious hatching under unfavorable environmental conditions. Using immunohistochemistry, pharmacological inhibition of matrix metalloproteinase 13 (Mmp13), and in vivo zymography, we show that Mmp13a is present in the hatching gland just as embryos become hatching competent and that Mmp13a activity is required for both normal hatching and hypoxia-induced precocious hatching. We conclude that Mmp13a likely functions in activating the hatching enzyme zymogen and that Mmp13a activity is necessary but not sufficient for hatching in zebrafish. This study highlights the broad nature of MMP function in development and provides a non-mammalian example of extra-embryonic processes mediated by MMP activity.

| S-EPMC7151336 | biostudies-literature

Cryopreservation of Anopheles stephensi embryos.

Project description:The ability to cryopreserve mosquitoes would revolutionize work on these vectors of major human infectious diseases by conserving stocks, new isolates, lab-bred strains, and transgenic lines that currently require continuous life cycle maintenance. Efforts over several decades to develop a method for cryopreservation have, until now, been fruitless: we describe here a method for the cryopreservation of Anopheles stephensi embryos yielding hatch rates of ~ 25%, stable for > 5 years. Hatched larvae developed into fertile, fecund adults and blood-fed females, produced fully viable second generation eggs, that could be infected with Plasmodium falciparum at high intensities. The key components of the cryopreservation method are: embryos at 15-30 min post oviposition, two incubation steps in 100% deuterated methanol at - 7 °C and - 14.5 °C, and rapid cooling. Eggs are recovered by rapid warming with concomitant dilution of cryoprotectant. Eggs of genetically modified A. stephensi and of A. gambiae were also successfully cryopreserved. This enabling methodology will allow long-term conservation of mosquitoes as well as acceleration of genetic studies and facilitation of mass storage of anopheline mosquitoes for release programs.

| S-EPMC8741979 | biostudies-literature

Regulation of zebrafish hatching by tetraspanin cd63.

Project description:Tetraspanins cause the clustering of membrane proteins into a level of organisation essential for cellular function. Given the importance and complicated nature of this mechanism, we attempted a novel approach to identify the function of a single component in a biologically relevant context. A morpholino knockdown strategy was used to investigate the role of cd63, a membrane protein associated with intracellular transport and a melanoma marker, in embryonic zebrafish. By using three separate morpholinos targeting cd63, we were able to identify a specific phenotype. Strikingly, morphant fish failed to hatch due to the lack of secreted proteolytic enzymes required for chorion-softening. The morphology of the hatching gland at both the cellular and intracellular levels was disorganised, suggesting a role for cd63 in the functioning of this organ. This work identifies a specific role for cd63 in the zebrafish embryo and provides evidence for the suitability of zebrafish as a model system for the investigation of tetraspanin enriched microdomains.

| S-EPMC3098263 | biostudies-literature

Triazole Fungicides Inhibit Zebrafish Hatching by Blocking the Secretory Function of Hatching Gland Cells.

Project description:In animals, hatching represents the transition point from a developing embryo to a free-living individual, the larva. This process is finely regulated by many endogenous and environmental factors and has been shown to be sensitive to a variety of chemical agents. It is commonly evaluated in bioassays in order to establish the effects of different agents on early development and reproductive capabilities in fish and other aquatic animals. In fish, the breakdown of the chorion is achieved by the secretion of choriolysin by hatching gland cells (HGCs) into the perivitelline space (PVS), coupled with spontaneous movements of the developing larva. In this work, we used zebrafish to assay the effects of a family of widely used agrochemicals-triazoles Triadimefon (FON), Triadimenol (NOL) and free triazole (1,2,4-T)-on hatching success. We found a strong inhibition of hatching by triazole exposure which was correlated with morphological changes and a reduction in the secretory function of the HGCs. As a consequence, the release of choriolytic enzymes by HGCs was reduced. We also found that HGC secretion reduction after exposure to FON can be rescued by co-incubation with a dopamine D2 receptor antagonist but not by antagonists of the D1-like receptors. This suggests a specific pathway through which this family of fungicides may be impairing a critical event in the fish life cycle.

| S-EPMC5412296 | biostudies-literature

Correlative Light and Electron Microscopy of Rare Cell Populations in Zebrafish Embryos Using Laser Marks.

Project description: Not available

| S-EPMC4677546 | biostudies-literature

Effect of laser-assisted hatching and necrotic blastomere removal on the development of vitrified-warmed four-cell mouse embryos.

Project description:PURPOSE: To investigate the effect of laser-assisted hatching and necrotic blastomere removal on the development of vitrified-warmed mouse embryos. METHODS: The vitrified-warmed four-cell stage mouse embryos were divided into five groups; vitrified intact with no laser-assisted hatching, vitrified intact with laser-assisted hatching, vitrified damaged with neither laser assisted hatching nor necrotic blastomere removal, vitrified damaged with laser-assisted hatching, and vitrified damaged with necrotic blastomere removal. Thereafter blastocyst formation, blastomere and apoptotic cell number within all groups were statistically compared. RESULTS: The rate of blastocyst formation showed a significant improvement in the group vitrified intact with laser-assisted hatching. However, neither laser-assisted hatching nor necrotic blastomere removal can improve a delayed vitrified-warmed damaged embryos in term of blastocyst formation and total cell number. Nevertheless, apoptotic cell number was significantly reduced after application of both techniques. CONCLUSIONS: Laser-assisted hatching can improve the development of vitrified-warmed intact four-cell stage mouse embryos, whereas necrotic blastomere removal has no significant effect on the development of vitrified-warmed four-cell stage damaged embryos.

| S-EPMC2596673 | biostudies-other

OmicsDI is part of the ELIXIR infrastructure

OmicsDI is an Elixir interoperability service. Learn more ›

Tweets

OmicsDI Databases

PRIDE
PeptideAtlas
MassIVE
JPOST Repository
Physiome Model Repository

EGA
EVA
ENA
LINCS
PAXDB
Cell Collective

MetaboLights
Metabolomics Workbench
MetabolomeExpress
GNPS
BioModels
FAIRDOMHub

ArrayExpress
dbGaP
ExpressionAtlas
GEO
NODE

Information

Databases
Help
API
Contact us
Code on GitHub
Terms of use
Submit Data