Cloned embryonic primary cells are manufactured and transplanted using the following procedures.

Embryll’s cloned embryonic primary cells are currently being provided to aging pets. Transplantation of cloned embryonic primary cells is performed by an Embryll accredited veterinarian. From the beginning stage of determining the type of cells to the final stage of transplanting cells, the entire process proceeds in the following order through close collaboration between an Embryll accredited veterinarian and Embryll scientists. The time to manufacture cells varies depending on the type of pet, but it usually takes three months.

STEP 1. Comprehensive medical examination

An aging pet receives a comprehensive medical examination at a nearby veterinary hospital. After the checkup, a veterinarian and the owner of the pet decide the types of cells to be transplanted into the pet.

STEP 2. Ordering cells

On the Embryll website, the veterinarian orders cells to be transplanted to the pet.

STEP 3. Sending a tissue collection kit

Embryll sends a tissue collection kit to the veterinary hospital with an insulated container to transport the kit at low temperatures.

STEP 4. Collecting tissues samples

The veterinarian collects small tissues around 5 mm in diameter from the pet’s skin and transfers them to a special preservation solution in the kit, which is stored in the insulated container.

STEP 5. Shipping the tissues

The veterinarian sends the insulated container promptly to the Embryll’s laboratory through FedEx Priority Overnight® on the same day the tissues were collected. During this process, the tissues are preserved at low temperatures.

STEP 6. Culturing the tissues

Embryll scientists culture the collected tissues of the pet, and healthy tissues are separately stored from them.

STEP 7. Retrieving oocytes

Oocytes are retrieved from young and healthy animals that are the same species as the pet. This step is the same process as a gynecologist retrieving oocytes from a woman for in vitro fertilization in a fertility clinic.

STEP 8. Removing DNA from the oocyte

A micropipette with a diameter of 10 um (1/10th the width of a human hair) is inserted into the oocyte, and DNA inside the oocyte is pulled out and removed. In animal cloning, this step is called enucleation. The oocyte can be damaged during enucleation, so this step should be carried out very carefully.

STEP 9. Cell fusion and cell reprogramming

A single cell is isolated from the healthy cultured tissues, pulled in a micropipette and inserted into the oocyte. The fusion of the oocyte and the cell is induced by sending currents lower than 100 volts for 20 us several times to media containing the oocyte. A zygote fused with the oocyte and the cell starts reprogramming.

STEP 10. Transfer and retrieval of the cloned embryo

When the zygote starts cell division and divides into two cells, this cloned embryo is transferred into the uterus of an ovulating surrogate mother, and the embryo is developed to the stage of organ formation. The cloned embryo is retrieved from the surrogate mother in a timely manner when primary cells can be manufactured from this embryo.

STEP 11. Manufacturing cells

High purity primary cells are manufactured from the retrieved cloned embryo by separating and purifying the tissues of each body part for transplantation. Quality control tests and genetic tests are performed for cell viability, cell purity, and the presence of contamination from microorganisms. Then, qualified cloned embryonic primary cells are stored in liquid nitrogen at -196°C.

STEP 12. Cell delivery

Embryll sends the cloned embryonic primary cells to a veterinary hospital through cold chain transport, keeping the cells frozen.

STEP 13. Cell transplantation

The veterinarian transplants the cloned embryonic primary cells into the pet. The aging pet begins to regain youth.

Through cell transplantation, the aging pet naturally reclaims its youth.

The manufacturing process of cloned embryonic primary cells is very similar to the process of cloning dogs or cats. If a cloned embryo in a surrogate mother’s uterus is left there without being retrieved, a cloned animal will be born. This way, the DNA of primary cells and pets is 100% identical, because the cloned embryonic primary cells are manufactured through animal cloning protocols. Therefore, the immune rejection problem, one of the biggest obstacles to cell therapy, can be solved fundamentally.

Additionally, cloned embryonic primary cells are simply physically dissociated cells from a cloned embryo and the manufacturing process does not involve artificial modifications. This has a very important meaning in predicting the in vivo safety of cells as a treatment. On the other hand, stem cells undergo several artificial modifications during the process of cell therapy manufacturing. For example, in the case of induced pluripotent stem cells, processing steps to reprogram somatic cells to stem cells involve using viruses, plasmid vectors, mRNA, or proteins, and those steps induce the overexpression of somatic cells’ four transcription factors (Oct4, Sox2, Klf4, C-Myc). Likewise, the process to transform embryonic stem cells into cell therapy involves several modifications such as differentiation and proliferation. However, artificially manufactured cells have latent risk factors such as genetic toxicity or chromosome mutations, so in vivo transplantation of artificially manufactured cells often causes side effects like tumors. For the aforementioned reasons, there is no existing FDA-approved stem cell therapy yet in the world. Conversely, cloned embryonic primary cells are simply physically dissociated and purified cells from a cloned embryo that has completed differentiating into all tissues and organs by itself. Manufacturing these cells does not involve any artificial modifications. It is so-called ‘Cell therapy manufactured by nature.’

Most importantly, the greatest advantage of cloned embryonic primary cells is their youth. An embryo is the earliest stage of life, and the tissues separated during this stage are full of young cells, which have healthy vitality and immense ability to divide. Aging pets will naturally restore their youth when these cells are transplanted into the pets.

Then, specifically which cells need to be transplanted to regain youth? Here is a detailed explanation.

References

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[3] de Munter JP, Wolters EC. Autologous stem cells in neurology: is there a future? J Neural Transm (Vienna). 2013 Jan;120(1):65-73. doi: 10.1007/s00702-012-0913-9. Epub 2012 Nov 23. PMID: 23180301.

[4] Takahashi K, Yamanaka S. Induction of pluripotent stem cells from mouse embryonic and adult fibroblast cultures by defined factors. Cell. 126 (4): 663–76. doi:10.1016/j.cell.2006.07.024. PMID 16904174.