- As we know recently Indian scientists have completed a milestone journey by cloning the world’s first buffalo named Samrupa. But, cloning is the most tedious process in animal biotechnology and there are several reasons for its success rate being very less. In the case of Dolly, the first sheep cloned 13 years ago, out of the 272 trials only five were successful and this time during samrupa’s cloning, out of 100s only one has become successful and that new born calf also died within 5 days.
Selection of defective cells
Genetically defective variety selection
Non expertise in cloning
Infection during cloning
Contamination of the media
Instruments used may be source of contamination
Maintaining suitable environment
Endotoxin, the main culprit
Selection of suitable surrogate mother
Post birth precaution
Following may be the 10 most important hurdles in the process of cloning
The animal to be cloned must be in a good health condition and disease free. Even the cells/tissues selected should be defect free both at nuclear and cytoplasmic levels.
If the host animal has some hereditary defects then chances of inheritance of the same defect into the offspring is more. So, always genetically improved variety of animals should be selected for cloning.
The scientist team involved in the cloning process must be experts in the cloning process although the success rate for cloning is less than 1 % of all samples taken.
There are always chances of bacterial and viral infection to the samples during cloning.
After enucleation of theÂ host cell and fusion with surrogate mother’s egg cells the recombined cells are grown on artificial media upto blastular eight cell structure. The media used here are most susceptible source of contamination.
Instruments like knives, blades, needles, sonicators , dishes and others must be properly cleaned and sterilized before use.
Incubators, laminar air flows and lab environment are itself sources of contamination. So, aseptic condition and level IV safety measures should be practiced during cloning.
Even after sterilization, the dead Gram negative bacterial cell wall components (called endotoxins) are the main source of infection. These are of nano grams size and lipopolysaccharide in nature and stable in heat and chemical conditions. So, the cloning process and materials must be protected from endotoxins contaminations.
The mother whose ovum and Uterus will be used for the growth of foetus should be disease free and genetically improved.
Even after the birth of the offspring contamination free environment should be maintained for a few days or even for month. Since newly born organisms are more susceptible to infections. Like in the case of Samrupa, it died due to respiratory infection within five days of her birth.
Expertise, proper handling, selection of suitable trait and endotoxins free processing are required for successful cloning. Indian scientists find many more opportunities of such milestones in animal cloning for the benefit of mankind by garnering the benefits of better livestock.