Prevention of Cross-contamination in Animal cell cultures

Prevention of Cross-contamination in Animal cell cultures

The following are the practices that help avoid to cross-contamination in animal cell lines:

*        Absence of cross-contamination may be confirmed by DNA fingerprinting, iso-enzyme analysis or DNA profiling karyotype.

*        As much as possible, cell lines should be obtained through a cell bank that has performed the proper validation of the cell line, we should authenticate the cell lines on our own level. ( please read :-
Detection of Mycoplasma Contamination in Animal Cell Cultures.)

*        Rapidly growing lines like HeLa (human cervical cancer cell line) must be handled carefully and protected from other cultures.

*        Use of one pipette for different cell lines should be avoided.

*        We should not open culture flasks of different cell lines or media bottles at one time, they must me opened simultaneously as per requirement.

*        Always try to add medium or any other reagents to the flask as the first step, and then after that add the cells.

*        Do not use same bottle of medium or trypsin for different cell lines.

*        Never put a once used pipette back in the bottle containing medium or trypsin etc., after using it in a culture flask of cells.

*        All the characteristics of culture must be checked regularly and any suspected change in morphology, other phenotypic properties or growth rate.

*        Avoid using unsterilized or unplugged pipettes and tips, for preventing cross-contamination.

 

Occurrence of cross-contamination is mostly possible during the experiments. But, to avoid them, it is very important that all precautions should be followed and the checking of cell strain and its characteristics must be regular.

Also not to forget these rules for avoiding Cross Contamination:

·         Whenever possible, characterization of new cell lines must be done by DNA fingerprinting immediately after isolation.

·         Any new lines that are introduced into the lab must be quarantined unless we are sure that they are not contaminated.

·         For contamination due to mycoplasma, use fluorescent staining of fixed preparations or PCR; while for regular contaminations, use normal or phase contrast microscopy.

·         A culture should not be decontaminated if it is replaceable.

·         Replacement of cultures must be done carefully with proper handling.

·         Do not keep all cultures in antibiotics always. At least one set culture of each cell line should be maintained without using antibiotics for about two weeks minimum and could be continued to detect and identify any possible contaminations.

·         Never share solutions or media among cultures or cell lines; the cell line characters must be checked periodically to prevent cross-contamination.

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Did you read ? 
Maintenance of Sterility in Animal Tissue Culture.

Also read: 
Detection of Mycoplasma Contamination in Animal Cell Cultures.

Viral Contamination in Animal cell cultures.