Partnership. Performance. Success.

Despite news reports of breakthroughs in the cryopreservation of human egg cells (unfertilized ova) over the past seven years, there in fact remains no reliable, validated, and highly successful method for the preservation of human ova at cryogenic temperatures. But at 21CM, we’ve made significant cryobiological advances that are rapidly improving the prospects for making this important technology a reality.

We’re currently looking to partner with assisted reproductive technology centers, fertility clinics and endocrinologists to further our research in determining the ability of human ova to survive true vitrification using 21CM proprietary methodologies.

Advance the future of reproductive medicine; contact us toll-free at 866-889-1215.

The Benefits of Technology

To solve the inherent problems of conventional cryopreservation, we use a vitrification process, which allows us utilize ultra rapid cooling rates to “outrun” chilling injury without the development of intracellular ice. Vitrification as a cryopreservation method has many significant advantages and benefits, including the escape from damage stemming from both intracellular and extracellular ice formation.

The Benefits of 21CM Research

We provide contract research support for reproductive technology initiatives that have a need to identify preservation methods without causing damage to the mammalian ova. Our team consists of a dedicated project manager plus technical and executive account managers who facilitate each project's design, implementation, evaluation, and ultimate commercial deployment. We offer:

• Experienced multidisciplinary project teams
• Proactive communication and innovative solutions
• Responsiveness to sponsor's individual needs
• Extensive protocol reviews
• Consultative services, including project design assistance
• Coordination between service and product needs
• Customized solution development
• Access to our cutting-edge cryobiology laboratory
• Solution preparation, sterilization, packaging, and international shipping capabilities
• Device design and prototype fabrication capabilities

Filling the Reproductive Technology Gap

To solve the inherent problems of conventional cryopreservation, our scientific investigations center on a process called vitrification, a process that is able to utilize ultra rapid cooling rates to “outrun” chilling injury but without the development of intracellular ice. This process leads to the conversion of intracellular and extracellular liquid into an amorphous solid without the formation of crystals (i.e., to the formation of a glass, rather than ice). Vitrification as a cryopreservation method has many significant advantages and benefits, including the escape from damage stemming from both intracellular and extracellular ice formation.

This allows a significant increase in cooling rates, which permits processing times to be shortened, expensive controlled-rate freezing devices to be dispensed with, and fluctuations in outcome due to such uncontrolled variables as varying sample-to-sample nucleation temperatures to be eliminated. Additionally, we have developed direct methods of reducing chilling injury beyond the mere reduction in time for it to occur through the use of rapid cooling.

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Many variables in the vitrification process still exist that can profoundly influence its effectiveness and its’ potential to improve the survival rates of vitrified oocytes. These include, but are not limited, to:

1. Type and concentration of cryoprotectant (almost every kind of cryoprotectant has some degree of toxicity)
2. Temperature of the vitrification solution at exposure
3. Duration of exposure to the final cryoprotectant concentration
4. Type of container used for vitrification (this influences cooling rate)
5. Quality and developmental stage of the oocyte
6. Method used to add the cryoprotectant to the ova
7. Method used to remove the cryoprotectant from the ova

Embryos have been cryopreserved and stored successfully for many years. Embryos are eggs that have been fertilized, and are approximately the same size as mature eggs, but more likely to survive cryopreservation. This is due in part to the fact that their sensitivity to chilling injury is all but eliminated due to dramatic changes induced in the cell membrane by fertilization.

These changes allow embryos to be cooled more slowly and therefore to more easily escape intracellular ice formation. However, the cell shrinkage that occurs with extracellular freezing can also lead to cellular injury and death. Embryos also appear to be more able than ova to tolerate the physical and osmotic stresses associated with extracellular ice formation.

The zona pellucida is an extracellular “shell” that completely surrounds the egg. It undergoes changes at the time of fertilization to prevent multiple sperm from fertilizing the egg. Cryopreserved oocytes are fertilized by a technique called intracytoplasmic sperm injection. ICSI has led to higher success rates utilizing cryopreserved oocytes, but is an intensive and expensive procedure.

Our mission is to create a cryopreservation method that minimizes or totally eliminates damage to the zona pellucida and allows fertilization by safer and less expensive and intensive methods. While we believe that the superior performance and convenience of vitrification will drive the development of this technique to higher levels of clinical efficiency and utilization, much more work remains before this will be a routinely available and viable option for most people who could benefit from it.

We are pleased that our preliminary results with vitrified unfertilized mouse ova as an initial model system showed advanced embryonic development at 80% of the frequency obtained with untreated control ova after rewarming. This was particularly encouraging because in these experiments, fertilization after vitrification with mouse sperm was done without ICSI, and the development rate of 80% of control rate was obtained without subtracting any problems arising from defective fertilization after vitrification and rewarming.

These initial results were published early in 2004. See Improved Vitrification Solutions Based on the Predictability of Vitrification Solution Toxicity, Cryobiology, Volume 48, pages 22-35, 2004.