
How to Improve Data Productivity by Avoiding Cell Line Contamination - benniebio
https://www.creative-bioarray.com/Services/Short-Tandem-Repeat-Analysis.htm
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benniebio
How to Avoid Cell Line Contamination - Mycoplasma detection 1\. Mycoplasma
culture, which is regarded as a standard method of detection. This is the most
sensitive method, depending on the standard agar plate on the selective and
highly enriched growth medium. The appereance will be unique ‘Omelette’, the
positive results with this method can prove the mycoplasma contamination.
However, this method can not detect all the speices, such as Mycoplasma
hyorhinis, DNA and PCR analysis is needed to ensure there is no contamination
happened! 2\. DNA Staining: DNA staining relies on the Hoechst 33258 stain,
which causes DNA-rich nuclei and any mycoplasma in the cytoplasm to fluoresce.
The use of this method may lead to false positive for the contamination due to
cell apoptosis or debris, or false negatives as this is the least sensitive
method. 3\. PCR: PCR-based mycoplasma detection can be very sensitive and can
detect up to 20 copy copies of mycoplasma genomes in 2 μL of sample.
Mycoplasma detection was achieved by amplifying the highly conserved 16S rRNA
operon coding region of the mycoplasma genome. 4\. Cell-Line Authentication.
ICLAC, which co-built by ECACC, representatives from other internation cell
culture organisations and respectable scientist commit to support the
importance of cell line authentication, creating a searchable database of
cross-contaminated and misidentified cell lines. This can be used to detect
cell line contaminations with the regularly updated database. 5\. Short Tandem
Repeat Profiling. With ASN-0002-2011 standard method, which analyzed 13
specific sites of STR and several additional sites to determine the integrity
of human cell lines authenticity.

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benniebio
What will Cause Cell Line Contamination? Cell Line nature enemies include:
Bacteria, viruses, mycoplasma, yeast and other eukaryotic cells. in the
culture procedure, what we offered below will lead to cell line contamination:
A. Confusion of the Mark, which refers to paste the wrong label in the the
daily culture and the management of the cells. B. Cross Contamination of
Cells, cell line is contaminated by other cells due to cell reagent utilities,
reusable or simultaneous operation of a variety of cells and other reasons. If
exogenous cells have a faster propagation rate than the original cell line,
exogenous cells will gradually replace the original cell line. C. Cell culture
enviroment change will lead to cell line contamination. Besides, bacteria,
mycoplasma contamination or drug treatment may also change the type of cells.

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benniebio
What is Cell Line Contamination? Scientists considers the cell line is
contaminated while the cell line genotype is different from its original
donor. So for determining whether the cell line is contaminated usually
starting from the genotype. The contaminated cell lines can be classified into
the false cell line (Eukaryotic cell contamination occurs in the early stage
of the cell line, the original cells have ceased to exist, which has
completely become another cell line), and the misidentified cell line (
Eukaryotic cell contamination occurs after the cell line is established, and
the original cells and newly mixed cells are present in the cell line).

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benniebio
Why it is important for avoind cell line contaminations Cell line
contamination poses a serious threat to the integrity of biomedical research.
Following best practices for cell culture validation, storage and processing
can help address this ongoing challenge, thereby improving the reliability of
experimental research data.

