How often should you authenticate cell lines?

once every two months
How often should cell lines be authenticated? Cell lines should be tested: When a new cell line is established or acquired. Before cells are frozen, or once every two months while the culture is actively growing. At the start of a new series of experiments, and prior to publication.

Why is cell line authentication important?

Cell and microbial authentication is of the utmost importance; skipping the authentication step can lead to lost time and money and retractions of publication as cross-contamination or misidentification has invalidated the data that that research is built upon.

How do you establish a cell line?

The simplest way to create a new cell line is to modify an existing one, a common strategy when an established line already comes close to meeting the requirements. Cells optimized to grow particular viruses or maximize recombinant protein production often come from such modifications.

How is mycoplasma detected in cell culture?

Mycoplasma contamination can be detected by polymerase chain reaction (PCR). PCR is easy, sensitive, specific, fast, reliable, efficient and costeffective. The PCR test is based on the detection of 16S rRNA molecules of the most common species of mycoplasma contaminating cell cultures.

What are types of cell line?

Understanding Types of Cell Lines

Frog cell lines.
Hamster cell lines.
Mouse cell lines.
Rat cell lines.
Dog cell lines.

What is cell line and its types?

The term cell line refers to the propagation of culture after the first subculture. In other words, once the primary culture is sub-cultured, it becomes a cell line. A given cell line contains several cell lineages of either similar or distinct phenotypes.

How do you identify cell lines?

Cell lines can be tested and identified using multiallelic variable number of tandem repeats (VNTR). HLA typing and DNA fingerprinting using short tandem repeat (STR) and a variable number of tandem repeats for intra-species cross-contamination have been used for cell line identification.

How do I confirm my cell line?

Verification of the species of a cell line can be performed by probing the sequence associated with species-specific cytochrome c oxidase subunit 1 (CO1), commonly referred to as DNA barcoding [26,27], or by employing PCR assays with other species-specific primers [23].

What is cell line authentication?

Cell lines are fundamental research tools that serve as models for complex biological systems, especially those of human tissue. A widely used method for cell line authentication involves STR profiling, a PCR-based technique that compares multiple short tandem repeat (STR) markers between two or more cellular genomes.

How do you identify a cell line?

Are there methods to authenticate nonhuman cell lines?

Methods to authenticate nonhuman cell lines are lacking. Our efforts have focused on developing methods for authentication of important nonhuman cell lines to achieve measurement assurance of their use.

Why mouse cell line STR assays?

Development of Mouse Cell Line STR Multiplex Assays and a Consortium for Validation of the Measurement Method Mice have been the most popular laboratory animal, widely used for genetic and basic research studies, resulting in mouse cell lines being the most numerous and important cell lines second to human cell lines.

Is there a patent for STR marker authentication?

U.S. patent (No. 9,556,482 ) for an authentication method using NIST-identified short tandem repeat (STR) markers. U.S. patent application ( 20170101677) published for additional STR markers for the mouse cell line authentication method. NCBI BioSample Database entry for mouse STR profiles.

How are STR markers used to identify human cell lines?

The STR markers are highly polymorphic (many differences in repeat length) and are used to fingerprint the cell lines for authentication. We first developed a multiplex PCR assay to authenticate African green monkey cell lines and determine contamination by human cell lines (BMC Biotechnology 2011, 11:102).