Frequently Asked Questions

1. What are the advantages of the dry culture media?

Shipping costs are lower because refrigeration is not required during transport. Dryform cobfersculture media with a have a much longer shelf life compared to their liquid counterparts, allowing storage for at least 1 year at 4°C. They can also be stored at room temperature for at least 5 months.

Additionally, the absence of water makes the culture media less susceptible to microbiological contamination.

2. How do I reconstitute dry culture media into liquid form?

EmbryoCloud provides the necessary amount of ultrapure water for each dry culture medium. Simply add the indicated amount of water to the vial containing the powder.

3. How long should I wait before using the culture medium after adding ultrapure water?

EmbryoCloud recommends dissolving the powder for at least 6 hours at a warm temperature, ideally overnight. Alternatively, the powder can be dissolved at 4ºC in the refrigerator for a minimum of 8 hours.

4. What should I do if the culture medium hasn’t dissolved properly?

EmbryoCloud recommends vortexing the resuspended powder for 10 seconds and ensuring there are no suspended particles remaining.

5. Is the culture medium ready to use once dissolved?

Yes, once reconstituted the medium is ready to use. However, EmbryoCloud recommends filtering the medium before use with a 0.22 µm pore size filter. Additionally, the maturation, fertilization and embryo culture media should be equilibrated for at least 2 hours before use undera 5% CO₂ atmosphere.

6. Can I continue using my routine protocol for in vitro embryo production?

Yes, absolutely. Once reconstituted, the medium can be used like any other liquid culture medium.

7. Do I have to use the entire volume of the culture medium, or can I save some for later?

Use only the amount you need. The remaining medium can be stored at 4°C, tightly sealed, for up to 1 month without compromising results.

8. What quality controls are performed on the dry culture media?

Each batch of culture medium is tested for pH, osmolarity, endotoxins, bacterial, and fungal contamination. Additionally, bovine embryonic assays

(BEA) or porcine embryonic assays (PEA) are conducted. These involve producing in vitro embryos from slaughterhouse ovaries and frozen semen.

Embryonic assays are evaluated based on the cleavage rate (percentage of zygotes divided 24 hours post-fertilization) and the blastocyst rate on days 7 and 8 post-fertilization (percentage of blastocysts relative to the total number of oocytes).

9. What results can be expected for embryo production with these media?

In EmbryoCloud’s embryonic assays using slaughterhouse ovaries and frozen semen, cleavage rates exceed 70%, and blastocyst rates exceed 20% on day 7 post-fertilization for both bovine and porcine species.

These are our quality control thresholds. However, due to biological variability (e.g., donor age, slaughterhouse origin, semen quality), results may vary, reaching up to 90% cleavage and 45% blastocyst rates for bovine and porcine oocytes.

10. For which animal species are these media designed?

EmbryoCloud offers a complete range of media for in vitro embryo production in both bovine and porcine species.

11. Do the culture media contain reproductive fluids?

Yes, the dry in vitro maturation media forthe NaturARTS line include follicular fluid so the composition of the culture media closer to the physiological conditions that embryos experience in vivo. Both in bovine and porcine species, results show better embryo production when oocytes are matured with media containing follicular fluid.

Reproductive fluids are tested for various pathogens before being used as supplements in the media and only fluids free of viruses are used:

• Bovine viruses: IBR, BVD, and bluetongue.
• Porcine viruses: Circovirus type 2, PRRS 1, PRRS 2, parvovirus. Additionally, they are free from other notifiable diseases:
• Bovine: Brucellosis, Tuberculosis.
• Porcine: African Swine Fever, Classical Swine Fever.

12. Are there alternatives to media containing reproductive fluids?

Yes, chemically defined media for in vitro maturation of oocytes where follicular fluid us replaced by BSA are available. However, our recommendation is using the NaturARTS line for the best results.

13. Are there publications using EmbryoCloud’s media or reproductive fluids?

Yes, these are available on our website (https://embryocloud.com/) or can be sent to you via email.