Collection of Tissue Samples

Pets are as close and important as the owner's family, but their lifespan is far shorter than that of humans, so pet cloning technology came into being in recent years. For many pet owners, cloning pets is considered to be the continuation of the life and emotion of previous pets. This technology seems to give pet owners a chance to reunite with their pets to make up for the pain and regret caused by the loss.

Theoretically, there are two procedures for artificial cloning of higher organisms in principle: one is embryo division, and the other is by transferring the nucleus to the egg or embryonic cells whose genetic material has been removed (nuclear transfer). Pet cloning discussed here refers to somatic cell nuclear transfer cloning technology. Specifically, clonal nuclear transfer technology is a technology to transfer genetic programs (nuclei with required genetic material) from totipotent mother cells or cells that are no longer totipotent (embryos, fetuses and even differentiated somatic cells) to unfertilized oocytes whose nuclei have been removed before. This technology essentially provides the possibility of replicating adult individuals and their genetic programs. The result is a new individual whose existence is not produced by fertilizing eggs by sperm cells.

Pet cloning is an extremely complex experimental process. The first step is to collect tissue samples from the target animals.

Operating Flow of Tissue Sample Collection

First, before collecting tissue samples, the experimenter designs the experimental procedure in advance, prepares the experimental materials, surgical instruments, protective measures, tissue freezing and transportation equipment, and keeps the surgical instruments, containers and other materials clean throughout the process to avoid contamination. In addition, the whole process of sample collection, including sample transportation, needs to be strictly kept at low temperatures to avoid repeated freezing and thawing of tissue samples.

The second step is to formally cut tissue samples from the target animals. Due to the rapid metabolism of metabolites in the body, sampling is a complex process. Both sampling and pretreatment need to be completed quickly and controlled within the same time range, and the sampling sites must be consistent.

In the third step, in order to avoid blood contamination, use deionized water or isotonic solution such as normal saline to clean the residual blood, and absorb the surface liquid with filter paper to obtain clean sample materials. Large pieces of tissue need to be cut into small pieces. The tissue sample should be cut into small pieces of about 20 ~ 50mg on ice. Of course, the size of the small piece can be adjusted appropriately according to the type of tissue sample.

The fourth step is to use aluminum foil paper to wrap tissue blocks and quickly freeze them in liquid nitrogen for about 10 seconds. The time of rapid freezing can be adjusted according to the size of the tissue block. It should be noted that the tissue should be frozen within 30 minutes after resection and treatment.

In the last step, put the quick-frozen tissue block into the freezing tube and make a clear and firm sample number to prevent loss of label during freezing. The frozen tissue is transported and stored in liquid nitrogen or ultra-low temperature refrigerator.

Operation Suggestions And Precautions In Real Life

Effective sampling is the premise and key step in the process of pet cloning. Only after obtaining effective samples can researchers bring your pet back to life. So, what should you do if you lose your pet? Taking pet dogs as an example, we first need active skin cells as cloning materials. You can choose to take tissue samples from your dog. If your dog is still alive, cut a small piece of skin; if your dog is dead, you need to wrap it all over with a wet towel, and then put it in the refrigerator to keep it cold. Of course, if you are worried that you are not professional enough or the required materials cannot be prepared completely, pet owners can also choose to let professional company technicians collect tissue samples for their dogs. In this regard, BioVenic has a full range of relevant services. Experiments such as tissue sample collection, preservation, transportation and subsequent tissue separation can be completed in BioVenic.

References

1. Albl B, et al. Tissue Sampling Guides for Porcine Biomedical Models. Toxicol Pathol. 2016 Apr;44(3):414-20.
2. Cyranoski D. Cloning comeback. Nature. 2014 Jan 23;505(7484):468-71.
3. Loi P, et al. A New, Dynamic Era for Somatic Cell Nuclear Transfer? Trends Biotechnol. 2016 Oct;34(10):791-797.