Preservation of Embryonic Stem Cells

He, Xiaoming

doi:10.5772/13860

Cited by 9 publications

(12 citation statements)

References 153 publications

(185 reference statements)

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“…Such an ice-seeding step is not used for vitrification and the thermodynamics of vitrification is typically non-equilibrium. As a result, for the cryomicroscopy study of IIF during low-CPA vitrification, ice crystals typically form in the extracellular solution at much lower temperatures (approximately −20 °C, Figures 2 and S2) than the thermodynamic equilibrium melting point (usually higher than −10 °C [2b] ) of the solutions and they are usually very fine as their growth is hampered by the slow diffusion of water molecules ( i.e. , diffusion-limited growth) at such deep subzero temperatures.…”

Section: Discussionmentioning

confidence: 99%

“…[1] Because continuous expansion of cells in vitro by culturing at 37 °C is expensive and time-consuming and may result in uncontrolled spontaneous differentiation of stem cells, banking living cells for future use by cryopreservation is an enabling technology to the eventual success of the burgeoning cell-based medicine. [2] Cell cryopreservation is achieved by first cooling cells to usually below −80 °C so that all the biophysical and biochemical activities in cells are arrested ( i.e. , the cells enter a state of suspended animation), followed by warming back for use at a desired future time.…”

Section: Introductionmentioning

confidence: 99%

“…For the former, cells are gradually dehydrated due to exosmosis driven by the elevated osmolality of extracellular solution when extracellular water transforms into ice during freezing at a slow cooling rate (typically less than 5 °C min −1 ), which is referred as freeze concentration and can cause significant damages to cells. [2b,5] On the other hand, conventional high-CPA vitrification is achieved by using penetrating cryoprotectants (CPAs) such as dimethylsulfoxide (DMSO) and 1,2-propanediol (PROH) at concentrations that are much higher than that used for slow freezing (up to 8 M versus ~1–2 M), which is toxic to cells due to the high CPA-induced metabolic and osmotic (including cell dehydration) injuries. [6] As a result, multiple steps are required to gradually load (before cooling) and unload (after warming) the CPAs to potentially reduce the high-CPA cytotoxicity, which is time-consuming and stressful.…”

Section: Introductionmentioning

confidence: 99%

“…[6] As a result, multiple steps are required to gradually load (before cooling) and unload (after warming) the CPAs to potentially reduce the high-CPA cytotoxicity, which is time-consuming and stressful. [4b,7] To overcome these problems, low-CPA vitrification has been explored for cell cryopreservation by using various methods and devices such as electron microscopy grid and quartz microcapillary (QMC) [2a,4b,7d,8] to achieve ultra-rapid cooling rates that minimize IIF by reducing the time available for ice nucleation and growth. Although the cooling rates can be very high (up to ~10 6 °C min −1 ) to achieve vitrification (defined as no visible ice formation in this study in accordance with the literature [9] ) by plunging the devices into liquid nitrogen due to the boiling of liquid nitrogen, the warming rate by plunging the devices into an aqueous solution is usually much lower.…”

Section: Introductionmentioning

confidence: 99%

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Alginate Hydrogel Microencapsulation Inhibits Devitrification and Enables Large‐Volume Low‐CPA Cell Vitrification

Huang

Choi

Rao

et al. 2015

Adv Funct Materials

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133

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Cryopreservation of stem cells is important to meet their ever-increasing demand by the burgeoning cell-based medicine. The conventional slow freezing for stem cell cryopreservation suffers from inevitable cell injury associated with ice formation and the vitrification (i.e., no visible ice formation) approach is emerging as a new strategy for cell cryopreservation. A major challenge to cell vitrification is intracellular ice formation (IIF, a lethal event to cells) induced by devitrification (i.e., formation of visible ice in previously vitrified solution) during warming the vitrified cells at cryogenic temperature back to super-zero temperatures. Consequently, high and toxic concentrations of penetrating cryoprotectants (i.e., high CPAs, up to ~8 M) and/or limited sample volumes (up to ~2.5 μl) have been used to minimize IIF during vitrification. We reveal that alginate hydrogel microencapsulation can effectively inhibit devitrification during warming. Our data show that if ice formation were minimized during cooling, IIF is negligible in alginate hydrogel-microencapsulated cells during the entire cooling and warming procedure of vitrification. This enables vitrification of pluripotent and multipotent stem cells with up to ~4 times lower concentration of penetrating CPAs (up to 2 M, low CPA) in up to ~100 times larger sample volume (up to ~250 μl, large volume).

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

Alginate Hydrogel Microencapsulation Inhibits Devitrification and Enables Large‐Volume Low‐CPA Cell Vitrification

Huang

Choi

Rao

et al. 2015

Adv Funct Materials

Self Cite

133

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“…More recently, low-CPA vitrification has been explored for cell cryopreservation by using various methods or devices [8-10,12,16,34,37,39], to achieve ultra-rapid cooling rate that minimizes IIF by reducing the time available for ice nucleation and growth. For example, the miniaturized quartz microcapillary (QMC) has been successfully used to achieve low-CPA vitrification of mouse mesenchymal stem cells (MSCs), mouse embryonic stem cells (ESCs), and mouse oocytes [12,16,39].…”

Section: Introductionmentioning

confidence: 99%

Improved low-CPA vitrification of mouse oocytes using quartz microcapillary

Choi

Huang

2015

Cryobiology

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Cryopreservation by low-cryoprotectant (CPA) vitrification has the potential to combine all the advantages of the conventional high-CPA vitrification and slow-freezing approaches while avoiding their drawbacks. However, current low-CPA vitrification protocol for cryopreservation of oocytes requires a lengthy and multi-step procedure for unloading CPAs. In this study, we report a much-simplified procedure of using quartz microcapillary (QMC) for low-CPA vitrification of mouse oocytes with only one step for unloading CPAs. The immediate viability of oocytes after the improved low-CPA vitrification was determined to be more than 90%. Moreover, no significant difference was observed in terms of embryonic development from the two-cell to blastocyst stages between the fresh and vitrified oocytes after in vitro fertilization (IVF). This improved low-CPA vitrification technology has the potential for efficient cryopreservation of oocytes to preserve the fertility of mammals including humans for assisted reproductive medicine, maintenance of animal resource and endangered species, and livestock management.

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Cryopreservation: A Comprehensive Overview, Challenges, and Future Perspectives

et al. 2023

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Cryopreservation is the most prevalent method of long‐term cell preservation. Effective cell cryopreservation depends on freezing, adequate storage, and correct thawing techniques. Recent advances in cryopreservation techniques minimize the cellular damage which occurs while processing samples. This article focuses on the fundamentals of cryopreservation techniques and how they can be implemented in a variety of clinical settings. The article presents a brief description of each of the standard cryopreservation procedures, such as slow freezing and vitrification. Alongside that, the membrane permeating and nonpermeating cryoprotectants are briefly discussed, along with current advancements in the field of cryopreservation and variables influencing the cryopreservation process. The diminution of cryoinjury incurred by the cell via the resuscitation process will also be highlighted. In the end application of cryopreservation techniques in many fields, with a special emphasis on stem cell preservation techniques and current advancements presented. Furthermore, the challenges while implementing cryopreservation and the futuristic scope of the fields are illustrated herein. The content of this review sheds light on various ways to enhance the output of the cell preservation process and minimize cryoinjury while improving cell revival.

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Preservation of Embryonic Stem Cells

Cited by 9 publications

References 153 publications

Alginate Hydrogel Microencapsulation Inhibits Devitrification and Enables Large‐Volume Low‐CPA Cell Vitrification

Alginate Hydrogel Microencapsulation Inhibits Devitrification and Enables Large‐Volume Low‐CPA Cell Vitrification

Improved low-CPA vitrification of mouse oocytes using quartz microcapillary

Cryopreservation: A Comprehensive Overview, Challenges, and Future Perspectives

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