Use of Antibiotics in Animal Tissue Culture for maintaining sterility

Sterilization of Explant Tissues and Primary Culture by Antibiotics

Antibiotics are the compounds that either kill or inhibit the growth of bacteria and few fungi and protozoans. These can be used to avoid bacterial contamination in the cell culture.

Following are the steps for sterilization of tissues.

1. For example, the tissue like the human skin is collected and readily preserved in balanced salt solution or a liquid nutrient solution.
2. The tissue is then kept dipped in 70% isopropanol for 15-30 seconds.
3. The tissue is then removed from isopropanol and then dipped in Dulbecco’s Phosphate Buffer Saline (DPBS) solution for few seconds.
4. Tissue is removed from DPBS solution and then placed in another flask containing Eagle’s medium with Hank’s Balanced Salt Solution (HBSS) and antibiotics like penicillin, streptomycin and amphotericin.  The flask is then tightly closed and kept in refrigerator with temperature set at 4-5°C.
5. For subculture, the tissue is transferred into a petridish containing DPBS solution and swirled with a sterile forceps and then it is again transferred to another petridish containing DPBS solution and washed properly to remove blood and debris from tissue.
6. The tissue is cut into very small pieces of 1mm with sharp sterile instruments like scalpels, fine-tipped forceps and fine surgical scissors.
7. The solution is centrifuged to separate the tissue segments in form of pellet.
8. These pieces of tissue are then aseptically transferred into a tube containing DPBS solution and the tube is capped tightly and stored for further use.

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Animal Biotechnology - Tissue Culture and Cell Culture
Prevention of Cross-contamination in Animal cell cultures 
Maintenanceof Sterility in Animal Tissue Culture