Some Factors Affecting Embryo Storage in Laboratory Animals

Dg, Whittingham

doi:10.1002/9780470720332.ch6

Cited by 23 publications

(4 citation statements)

References 40 publications

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“…29 For example, a critical step for successful cryopreservation of embryos and oocytes for in vitro fertilization is control of ice nucleation whereby “seeding” is achieved by generating a cold spot on the outside of the cryocontainer. 30 Active ice nucleation is also a critical strategy for freeze-tolerant animals, 10,27 although ice nucleating agents (INAs) in the blood and gut/skin induce controlled freezing of extracellular water at multiple nucleation sites. 31 As the hemolymph gradually freezes, it is accompanied by an increase in the osmolality of the extracellular fluid, resulting in cellular dehydration as water is pulled from the intracellular environment.…”

Section: Discussionmentioning

confidence: 99%

“…It is generally accepted that the minimization of cryoinjury may be achieved when ice nucleation occurs as near as possible to the equilibrium freezing point . For example, a critical step for successful cryopreservation of embryos and oocytes for in vitro fertilization is control of ice nucleation whereby “seeding” is achieved by generating a cold spot on the outside of the cryocontainer . Active ice nucleation is also a critical strategy for freeze-tolerant animals, , although ice nucleating agents (INAs) in the blood and gut/skin induce controlled freezing of extracellular water at multiple nucleation sites .…”

Section: Discussionmentioning

confidence: 99%

See 1 more Smart Citation

Effect of Ice Nucleation and Cryoprotectants during High Subzero-Preservation in Endothelialized Microchannels

Tessier

Weng

Moyo

et al. 2018

ACS Biomater. Sci. Eng.

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Cryopreservation is of significance in areas including tissue engineering, regenerative medicine, and organ transplantation. We investigated endothelial cell attachment and membrane integrity in a microvasculature model at high subzero temperatures in the presence of extracellular ice. The results show that in the presence of heterogeneous extracellular ice formation induced by ice nucleating bacteria, endothelial cells showed improved attachment at temperature minimums of −6 °C. However, as temperatures decreased below −6 °C, endothelial cells required additional cryoprotectants. The glucose analog, 3-O-methyl-D-glucose (3-OMG), rescued cell attachment optimally at 100 mM (cells/lane was 34, as compared to 36 for controls), while 2% and 5% polyethylene glycol (PEG) were equally effective at −10 °C (88% and 86.4% intact membranes). Finally, endothelialized microchannels were stored for 72 h at −10 °C in a preservation solution consisting of the University of Wisconsin (UW) solution, Snomax, 3-OMG, PEG, glycerol, and trehalose, whereby cell attachment was not significantly different from unfrozen controls, although membrane integrity was compromised. These findings enrich our knowledge about the direct impact of extracellular ice on endothelial cells. Specifically, we show that, by controlling the ice nucleation temperature and uniformity, we can preserve cell attachment and membrane integrity. Further, we demonstrate the strength of leveraging endothelialized microchannels to fuel discoveries in cryopreservation of thick tissues and solid organs.

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Effect of Ice Nucleation and Cryoprotectants during High Subzero-Preservation in Endothelialized Microchannels

Tessier

Weng

Moyo

et al. 2018

ACS Biomater. Sci. Eng.

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“…Ice nucleation is believed to affect the cell structural, osmotic and colligative properties as well as is capable of causing ruptures and mechanical injuries through same means [5]. "Seeding" refers to one such method wherein an external preformed ice crystal is deliberately introduced as a nucleator into the system at a particular temperature [7,64,65]. A cold spot is generated generally outside of the container to minimize contamination.…”

Section: Controlling Ice Nucleationmentioning

confidence: 99%

Biophysics of Cryopreservation

SHARMA

2022

International Journal of Thermodynamics

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A significant credit towards today's scientific and medical advancements goes to the technique of cryopreservation. Cryopreservation refers to the maintenance of cellular life at subzero temperatures for a definite period of time in a state of suspended cellular metabolism. The technique has become an indispensable step in most scientific research and medical applications like assisted reproduction, transplantations, and cell-based therapies where-in it allows the long-term preservation of biological specimens like gametes, embryos, viruses, cells and tissues. Although already an extensively used technique, a significant proportion of the cryopreserved samples still incur notable damage. Ultimately this leads to a decreased post-thaw viability and proliferation. Moreover, it is also possible that events during the freezing process, provoke more serious disturbances in the preserved material with regard to its identity and functionality. Hence, with the need to use the technique more judiciously, additional studies are needed for optimizing the current cryopreservation methods in use. For this, a thorough understanding of the normal physiological changes that the cryopreserved sample undergoes and the physics of cryopreservation seems plausible. The review thus aims to unravel the current knowledge on the complex physico-chemical processes and reactions that occur during the standard cryopreservation techniques.

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“…Impurities, such as dust, act as ice nucleators, initiating large crystal formation at temperatures well above –39!C [18]. However, allowing ice crystals to form at temperatures more than a few degrees below zero during cooling has been shown to damage embryos and oocytes [21]. To initiate crystal formation in a solution at higher temperatures, the solution must be seeded with ice either by the addition of an ice crystal or by touching it with something colder, such as a forceps dipped in liquid nitrogen, as is done in the laboratory.…”

Section: Cryopreservation Techniquesmentioning

confidence: 99%