SHIRLEY – AUG 30, 2017 – Creative Bioarray, an innovative biotechnology company which concentrates on developing unique technologies for products and service, including professional cell line authentication that will help verify the identity of human cell lines, ensuring uniqueness of the cell line and detecting laboratory errors such as misidentification and cross-contamination of lines.
The first human cancer cell line was established in 1952. However, researchers found that many of the cultured cells had different characteristics from the original cell lines in later experiments, and that some methods of cell culture could lead to cell changes or cross-contamination. Cell culture system is very important in the research and development of bio-pharmaceuticals, the appropriate cell line authentication process gets a lot of attention from researchers. However, cell cross-contamination still is inevitable which affects every area of scientific research – from the bench to clinical trials. 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.
Microbial contamination of cell lines poses a serious challenge to obtaining reliable research data. The most dangerous microbial contaminants are mycoplasma. It can induce abnormal behavior of cultured cells, including altered growth rates, morphological changes, chromosomal aberrations and altered cellular metabolism. Contamination is difficult to control because mycoplasma lacks cell walls and therefore can not be treated with most antibiotics.
Mycoplasma detection is regarded as an effective method for avoiding cross-contamination. Low levels of mycoplasma contamination are difficult to detect and may require two or more detection methods.
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 appearance will be unique ‘Omelette’, the positive results with this method can prove the mycoplasma contamination. However, this method can not detect all the species, 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 2μL 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 international cell culture organizations 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 contamination with the regularly updated database.
5. Short Tandem Repeat Profiling. The human genome is full of repeated DNA sequences which come in various sizes and are classified according to the length of the core repeat units, the number of contiguous repeat units, and/or the overall length of the repeat region. DNA regions with short repeat units (usually 2-6 bp in length) are called Short Tandem Repeats (STR). STR gene locus consists of short tandem repeats of 3 to 7 base pairs, which are widely found in the human genome and can be used as highly polymorphic markers, that is DNA fingerprints of cells. The alleles on the STR gene locus can be distinguished by the number of copies of the repeats in the PCR (polymerase chain reaction) amplification region, which can be identified by fluorescence detection after capillary electrophoresis separation, followed by a certain calculation. According to the results obtained by STR classification and professional cell STR database comparison to calculate the sample belongs to the cell line or possible cross-contamination of the cell line name.
6. Reasonable storage and processing method will decrease the cell line contamination: Proper storage and handling of cells and cell culture reagents, as well as good sterile cell culture techniques, can minimize contaminations and improve reproducibility of experimental data.
About Cell Line Authentication offered by Creative Bioarray:
Creative Bioarray provides the world’s most comprehensive list of cells, and has realized that animal and human primary cells, tumor cell lines, continuous (immortalized) cell lines, and tissues are very important to the biopharmaceutical industry and to biomedical research as reagents, therapeutic modalities, and as proxy materials. Cell cultures have been at risk for misidentification due to labeling errors, incorrect classification by pathologists, and cross-contamination with other cell types. The sensitivity and high power of discrimination makes our STR analysis an ideal choice for the various types of cell authentication.
Company Name: Creative Bioarray
Contact Person: Chelsea Clark
Email: Send Email
Address:45-1 Ramsey Road Shirley
City: Long Island City
State: New York
Country: United States