The Journey of Jimmy - The Cryo Calf

This is sweet little Jimmy a year-old calf.  He was born on 2001 in Texas. He is a healthy calf who loves to play with his tail. There is something special about Jimmy. He is not naturally born. Once upon a time, a male bull named Brad died at the farm. Before he died, the owner farm Mr Clinton retrieved some of his samples of sperm. Brad had never successfully bred in his lifetime. Due to the good genetic, Brad’s sperm sample were stored and preserved by using cryopreservation method. Jimmy is the product from Brad’s frozen sperm.

Cryopreservation is process by which any living cell, tissue, organ or entire bodies are protected from decay by storing them at extremely low temperature. This method of preservation helps Brad’s sperm protected from decay by storing them at extremely low temperature. His sperm was stored at very low temperature which is at -195°C in CryoBioBank. At low enough temperature, any enzymatic or chemical activity which might cause damage to Brad’s sperm is effectively stopped.  

There are several types of method cryopreservation such as slow programmable freezing and vitrification (Lecchi et al., 2016). Brad’s sperm was cryopreserved by using vitrification method. Semen from all mammalian livestock species as well as poultry species can be successfully frozen. Freezing procedure for semen cryopreservation are species-specific, but the general procedures are as follows:

• Following collection, semen is diluted in a suitable ionic (salt) or non-ionic (sugar) solution adjusted to near physiological osmolarity.
• Suitable cryoprotectant is added, glycerol is mostly used, but dimethyl sulfoxide (DMSO), dimethylacetamide (DMA) or dimethylformamide (DMF) are also used especially in avian species.
• Diluted semen is cooled, sampled and then frozen in liquid nitrogen (-196 °C).
• Individual semen doses are generally frozen in straws rather than pellets to guarantee optimal sanitary conditions and permanent identification of each dose.

Mechanism of cryopreservation process

            Since the useful and successful of this method in preserving various type of foods, so there are many applications have been practiced in food industry. Cryopreservation protocols have been established for root and tubers, fruit, forest trees, ornamentals and plantation crops and often applied to large numbers of accessions within species.  Example of cryopreservation of pollen and seeds. Cryopreservation of pollen is preserving the pollen and it is useful for cross-pollination of cultivars differing in flowering period.

            In agricultural and horticultural cryopreservation of seeds, the species are tolerant to desiccation and exposure to liquid nitrogen. This method also as an alternative to the traditional storage. Celery is an example of orthodox seed cryopreservation. The number of genebanks and botanic gardens where cryopreservation is employed on large scale to different types materials is keep growing. Here some examples of cryopreserved collections of tropical plant species which are coffee, musa and cassava.

Cryopreservation is considered as sort of miracle in science world. Throughout the 1940s, researchers worked to determine a process for safely freezing cells that would be viable for use after thawing (Bailey, 2017). The first successful story of cryopreservation of mammalian cells was performed by Christopher Polge and his colleagues in 1949. They found that the frozen fowl spermatozoa had resumed motility on thawing and was capable of effecting fertilization on the egg, though no live chicks were produced. This had spark large interest amongst scientists on the potential of cryopreservation in cells preservation to produce living organisms.

          Subsequent studies resulted in key milestones to the enhancement of the technology. Smith and Polge (1950) had successfully cryopreserved bull spermatozoa; Stewart’s (1951) research resulted in the first live birth of calves after insemination using frozen spermatozoa; Latta (1971) involved in cryopreservation of plant cell cultures; both Whittingham and Wilmut (1972) had successfully recovered frozen mouse embryos; and Cohen and colleagues (1985) had stored human embryos using cryopreservation for use in in vitro fertilization. Thus, cryopreservation has become widely accepted as the optimal method for the preservation of microorganisms.

Recently, Malaysia had imported about RM5.5 billion of processed food, RM4.0 billion of sugar and sugar confectionary, RM3.0 billion of dairy products, RM1.8 billion of preserved vegetables and fruits and RM1.5 billion of cocoa (Mida.gov.my, 2017) due to lack source of food. Cryopreservation may assist to ensure the sustainability of food source by preserving the seeds of those species as they are unable to withstand dehydration process and sensitive to chilling (Engelmann & Dussert, 2013). As the source of food is sufficient, prices of food can be controlled during different season.

In addition, cryopreservation may assist in import export activity as it will guarantee a better quality of product. For example, conventional freezing of food may cause freezer burn, dehydration, colour changing, ice-crystallization, drip loss and lipid oxidation. Cryopreservation may assist to protect the physical and chemical properties of food. Transporting of food in long distance may affect the safety of food. This can be related to the experience where spices from India had been banned by certain country due to contamination and quality issue (Edison, 1995). As solution, extremely low temperature had been used during grinding to gain better quality of product and less risk of food contamination. Last but not least, every preservation methods may help in increasing the shelf life of product but in limiting time. Rapid cooling may assist to decrease the respiration rate of and retard the physiological changes of products thus increasing the storage life of food (Goswami, 2010).

As cryopreservation is the optimum method of preserving microorganisms, its potential has been studied and proven to be beneficial in food preservation. The technology of cattle breeding through insemination of frozen sperm by cryopreservation has been applied in the past 50 years ago and will continue so. Jimmy is not alone, he had, has and will be joined by his brothers and sisters through this technology.

References

Lecchi, L., Giovanelli, S., Gagliardi, B., Pezzali, I., Ratti, I., & Marconi, M. (2016). An update on methods for cryopreservation and thawing of hemopoietic stem cells. Transfusion and Apheresis Science, 54(3), 324–336. http://doi.org/10.1016/j.transci.2016.05.009  

Bailey, R. (2017, 5 October). “Revival of Spermatozoa after Dehydration and Vitrification at Low Temperatures” (1949), by Christopher Polge, Audrey Ursula Smith, and Alan Sterling Parkes. The Embryo Project Encyclopedia. Retrieved from http://embryo.asu.edu/pages/revival-spermatozoa-after-dehydration-and-vitrification-low-temperatures-1949-christopher

Polge, C., Smith, A. U. & Parkes, A. S. (1949). Revival of spermatozoa after vitrification and dehydration at low temperature. Nature 164, 666.

Smith , A. U. & Polge, C. (1950). Storage of bull spermatozoa at low temperatures. Vet. Rec. 62, 115-117.

Stewart, D. L. (1951). Storage of bull spermatozoa at low temperatures. Vet. Rec. 63, 65, 66.

Latta, R. (1971). Preservation of suspension cultures of plant cells by freezing. Can. J. Bot. 49, 1253, 1254.

Whittingham, D. G., Leibo, S. P. & Mazur, P. (1972). Survival of mouse embryos frozen to -196 and -296°C. Science 178, 411-414.

Wilmut, I. (1972). The effects of cooling rate, cryoprotectant agent and stage of development on survival of mouse embryos during freezing and thawing. Life Sci 11, 1071-1079.

Cohen, J., Simons, R., Fehilly, C. B., Fishel, S. B., Edwards, R. G., Hewitt, J., Rowland, G. F., Steptoe, P. C. & Webster, J. M. (1985). Birth after replacement of hatching blastocyst cryopreserved at expanded blastocyst stage.  Lancet I, 647.

Mida.gov.my. (2017). MIDA | Malaysian Investment Development Authority:. Food Technology and Sustainable Resources. [online] Available at: http://www.mida.gov.my/home/food-technology-and-sustainable-resources/posts/ [Accessed 19 Dec. 2017].

Engelmann, F., & Dussert, S. (2013). Cryopreservation. In M. N. Normah, H. F. Chin, & B. M. Reed (Eds.), Conservation of Tropical Plant Species (pp. 107-119). New York, NY: Springer New York.

Goswami, T. (2010). Role of Cryogenics in Food Processing and Preservation. International Journal of Food Engineering, 6(1).

Edison, S. (1995). Spices: Research Support to Productivity. Survey of Indian Agriculture.