iPS Cell Repository for Human Tissue and Disease Models
Innovation Showcase presented by ATCC at the ISSCR Annual Meeting, June 2011

ATCC has established an iPS Cell Repository to accession and globally distribute highly qualified, standardized cell lines that have been reprogrammed via expression of OSKM from a diverse range of tissues and disease states. Biorepositories accelerate research progress by managing the roadblocks often associated with the distribution of biological materials. View the video.

Introduction of Novel iPSC Disease Model and a Repository for the Preservation and Distribution of iPSC Lines
Noelle Strubczewski, Dezhong Yin, Yukari Tokuyama, Chengkang Zhang and Will Rust; ATCC, Manassas, VA.
Poster presented at the ISSCR Annual Meeting, June 2011

The ability to reprogram various types of somatic cells to generate human induced Pluripotent Stem Cells (iPSC) offers a new paradigm for modeling human tissue development and disease progression. These cells can also serve as a source for differentiated cells that can facilitate drug testing or enable cell therapy. However, the impact on these fields is largely determined by the breadth and genetic diversity of cell lines, and the accessibility of these lines to biomedical researchers around the world. The ATCC has established an integration-free reprogramming method using the Yamanaka factors delivered on episomal plasmids. We generate iPSC’s from primary human fibroblasts of healthy and diseased donors. Our primary fibroblasts are harvested from transplant and cadaveric organs with donor consent, or from the ATCC catalog, and are accompanied by donor medical history. Here we report the creation and characterization of a human iPSC from fibroblasts of a patient with a severe form of Osteogenesis imperfecta.

Respiratory virus infection of differentiated primary human small airway epithelial cells.
 W. Ullmer, J. K. Cooper, R. Marlow, B. Buck, T. Irish, ATCC, Manassas, VA.
Poster presented at the ASM Annual Meeting, May 2011

Background
The continuing emergence of viruses causing significant respiratory illness in humans highlights the need for in vitro models of the human airway epithelium to study virus biology and pathogenesis. In this study, we investigate the ability of ATCC® Primary Cell Solutions™ Human Small Airway Epithelial Cells (PCS-301-010) to differentiate into pseudostratified epithelium, as well as their susceptibility to infection by a panel of respiratory viruses, including isolates of the 2009 H1N1 influenza pandemic.

Methods
Cells were differentiated at an air-liquid interface for 28 days, with differentiation being determined by the expression of markers for cilia formation and secretory protein production. After 28 days of differentiation, the cells were infected with viruses, including 5 isolates of 2009 H1N1 influenza A virus, influenza B virus (B/Lee/40), respiratory syncytial virus (Long), parainfluenzavirus 3 (C 243), rhinovirus 16 (11757), adenovirus type 5 (Adenoid 75), and coronavirus (229E). Infection was monitored for 4 days, and resultant viral yields were determined by plaque assay for influenza viruses and 50% tissue culture infective dose (TCID₅₀) for all other viruses.

Results
Differentiation of the small airway epithelial cells was determined by quantitative reverse transcription polymerase chain reaction (qRT-PCR). The expression of genes involved in formation of cilia and Clara cell secretory protein (CCSP) were determined to be up-regulated during the differentiation process. We observed positive immunofluorescent staining for all of the viruses tested. Viral yields varied, but typically ranged between 10^2.5 and 10^5.5 IU/mL by 96 hours post-infection.

Conclusions
This work demonstrates the ability of primary human small airway epithelial cells to be used as an in vitro model that more closely represents the human small airway epithelium than standard cell culture. This model system may be a useful tool for the investigation of viral infections of the respiratory epithelium.

Rapid Identification of Cell Lines Sensitive to Clostridium perfringens Epsilon Toxin
By Matthew Boley, Kurt Langenbach and Brian Beck, BioServices Division, ATCC.
Poster presented at the ASCB Annual Meeting, December 2010

Abstract: The pore-forming epsilon toxin, produced by the ubiquitous anaerobe Clostridium perfringens, is a CDC/USDA overlap class B select agent and is of particular concern to the agricultural and biodefense communities. It is our hope that increased availability of human and animal epsilon-sensitive cell lines will facilitate research towards cellular receptor discovery and understanding trafficking mechanisms, subsequently strengthening the basis for toxin inhibition, treatment and countermeasure development. ATCC’s extensive cell biology collection was reviewed for promising epsilon-sensitive candidates, which were selected, then ranked according to target tissue, species origin and other criteria. Eighteen cell lines were chosen and screened for toxin susceptibility using the real-time, label-free, Roche xCELLigence™ System, which measures changes in impedance. Cell lines identified to be susceptible to the preliminary toxin challenge (100nM) were subsequently evaluated with a more expansive dosage range to determine EC₅₀’s and time to cytotoxicity. This work was performed using the xCELLigence System and a parallel battery of complimentary tests including a classical cytotoxicity assay, observation of cell morphology as well as the use of fluorescent cellular staining dyes to verify observed xCELLigence System and cytotoxicity data and to investigate observed blebbing and apparent syncytia formation. Twelve cell lines showed increased sensitivity with two cell lines exhibiting 8 and 20-fold sensitivity below the most sensitive cell line published to date. The identification of authenticated human and animal cell lines with differential responses to epsilon toxin provides a multifarious platform to investigate systems biology of receptor and cell-trafficking mechanisms for the purpose of cell-based toxin inhibition, treatment and countermeasure development.

ATCC Technology Assessment of Roche xCELLigence™ System — an
Electronic Impedance-Based Cell Sensing Unit
By Kurt J. Langenbach Ph.D., BioServices Division, ATCC, published online December 2010

Abstract: This article describes our systematic investigation into the utility of employing electronic impedance-based cell sensing measurement systems to evaluate changes in cell behavior. Our studies allowed us to evaluate multiple aspects of the system including sensitivity and flexibility. Results indicate that the xCELLigence System offers dynamic, real-time, label-free and non-invasive analysis of a variety of cellular events. In some scenarios, it offers considerable labor and reagent savings when compared to more classical approaches. In addition we found that the xCELLigence System offers an efficient way to optimize cell-based assays and provides data that is almost impossible to capture using typical endpoint assays.

Functional Cell Profiling of Endogenous GPCRs Using the xCELLigence™ System
By Jeff Irelan, ACEA Biosciences, Inc. and Jonathan H. Morgan, ATCC

Abstract: G-coupled protein receptors (GPCRs), also known as 7-transmembrane proteins, constitute the single largest class of therapeutic targets for clinical and investigational drugs. New technology for assaying receptor function in a cellular context has greatly increased the identification of novel regulators and reduced attrition rates (defined as failure in preclinical testing/clinical trials) for candidate compounds. The xCELLigence™ System from Roche Applied Science assesses real-time endogenous GPCR function in disease-relevant cells without using exogenous labels. Impedance-based real-time kinetic recordings can detect all second messenger mediated GPCR responses during the course of an experiment. In the present study, we show that the xCELLigence System is a sensitive and robust assay for continuously measuring endogenous GPCR function. A panel of 43 ligands encompassing 24 therapeutically relevant receptor families was examined, producing functional GPCR profiles for the commonly used cell lines, HeLa, U-2 OS, SH-SY5Y and CHO-K1 (ATCC® CCL-2™, HTB-962™, CRL-2266™ and CCL-61™, in that order), as well as the disease-relevant human primary cells: HUVECs (PCS-100-010) and mixed renal epithelial cells (PCS-400-012).

 Ending cell line contamination by cutting off researchers. Online article published August 08, 2010 at www.biotechniques.com. This article includes contributing quotes from Amanda Capes-Davies, a member of the ATCC Standards Development Organization Workgroup ASN-0002.
 
Abstract: After 50 years of skepticism, finger pointing and unenforced protocols, sentiments are growing for mandatory cell line authentication as a condition for funding and publication. The author investigates the current state of cell line contamination and finds how raising awareness could help cut off the supply of contaminated lines.

Recommendation of short tandem repeat profiling for authenticating human cell lines, stem cells, and tissues.  Authored by The ATCC Standards Development Organization Workgroup ASN-0002 and published online in the July 8, 2010 issue of In Vitro Cell. Dev. Biol. 

Abstract: Cell misidentification and cross-contamination have plagued biomedical research for as long as cells have been employed as research tools. Examples of misidentified cell lines continue to surface to this day. Efforts to eradicate the problem by raising awareness of the issue and by asking scientists voluntarily to take appropriate actions have not been successful. Unambiguous cell authentication is an essential step in the scientific process and should be an inherent consideration during peer review of papers submitted for publication or during review of grants submitted for funding. In order to facilitate proper identity testing, accurate, reliable, inexpensive, and standardized methods for authentication of cells and cell lines must be made available. To this end, an international team of scientists is, at this time, preparing a consensus standard on the authentication of human cells using short tandem repeat (STR) profiling. This standard, which will be submitted for review and approval as an American National Standard by the American National Standards Institute, will provide investigators guidance on the use of STR profiling for authenticating human cell lines. Such guidance will include methodological detail on the preparation of the DNA sample, the appropriate numbers and types of loci to be evaluated, and the interpretation and quality control of the results. Associated with the standard itself will be the establishment and maintenance of a public STR profile database under the auspices of the National Center for Biotechnology Information. The consensus standard is anticipated to be adopted by granting agencies and scientific journals as appropriate methodology for authenticating human cell lines, stem cells, and tissues.

Cell line misidentification: the beginning of the end.  Authored by The ATCC Standards Development Organization Workgroup ASN-0002 and published in the May 7, 2010 issue of Nature Reviews Cancer.

Abstract: Cell lines are used extensively in research and drug development as models of normal and cancer tissues. However, a substantial proportion of cell lines are mislabelled or replaced by cells derived from a different individual, tissue or species. The scientific community has failed to tackle this problem and consequently thousands of misleading and potentially erroneous papers have been published using cell lines that are incorrectly identified. Recent efforts to develop a standard for the authentication of human cell lines using short tandem repeat profiling is an important step to eradicate this problem.

What's in Your Vial?  Best practices for maintaining microbial QC strains.  Webinar presented by Elizabeth Kerrigan, Director, Standards and Certification, ATCC and Jaspreet Sidhu, Ph.D., Vice President of Business Development and Pharmaceutical Microbiology, Molecular Epidemiology, Inc.

Abstract: In the pharmaceutical and personal care industries, products, processes and environments are quality control tested to prevent microbial contamination. Microbial strains with confirmed identity, viability and purity, produced by meticulous laboratory procedures that minimize subculturing, are important components of quality control testing programs. Responding to industry demands for rapid microbiological testing, instrumentation and associated databases have been developed that allow for the standardized testing of phenotypic and genotypic traits across a wide array of microorganisms. A polyphasic approach to identification provides strain confirmation and avoids the pitfalls of misidentification, painful recalls and regulatory repercussions. Case studies will be provided to illustrate key points. (Hosted by BrightTALK™)

Impact of Microbial Contamination and Misidentified Cell Cultures on Research.  Webinar presented by Yvonne Reid, PhD, Cell Biology Collection Scientist, ATCC on March 9, 2010.

Abstract: Animal cell lines are important in vitro systems and tools for scientists in diverse disciplines beyond basic cell biology. Cell line authentication and characterization are crucial in these fields, yet most research scientists under appreciate them. Over the years, numerous cell lines have been shown to be misidentified due, in part, to poor techniques, inadequate authentication protocols, and sharing of unauthenticated cell lines amongst researchers. Technological advances have given rise to improved capabilities. Cell line authentication and characterization now require a comprehensive strategy that employs several complementary technologies for systematic testing for morphology, microbial contaminations, cellular cross-contamination as well as functionality. The validity of conclusions drawn from research data is dependent on consistent and unequivocal verification of cell line identity and function. It is estimated that the financial loss incurred by poorly characterized or misidentified cell lines is in the millions of dollars. An overview of the current technologies used to authenticate and characterize animal cell lines was presented. (Hosted by Corning)

Abstract: In the pharmaceutical and personal care industries, products, processes and environments are quality control tested to prevent microbial contamination. Microbial strains with confirmed identity, viability and purity, produced by meticulous laboratory procedures that minimize subculturing, are important components of quality control testing programs. Responding to industry demands for rapid microbiological testing, instrumentation and associated databases have been developed that allow for the standardized testing of phenotypic and genotypic traits across a wide array of microorganisms. A polyphasic approach to identification provides strain confirmation and avoids the pitfalls of misidentification, painful recalls and regulatory repercussions. Case studies will be provided to illustrate key points. (Hosted by BrightTALK™)

Good Cell Culture Practices.  Presented by Brian Douglass, Cell Biology Product Manager, ATCC.

Focus: Misidentification/cross-contamination, long-term subculturing and passage number, poor culture conditions, and microbial contamination (mycoplasma).

Corning Cell Culture Series. You are invited to a series of free web-based technical seminars on cell culture. Co-sponsored by ATCC and The Society for In Vitro Biology (SIVB), the webinars are designed to provide novel tips, best practices and proven techniques to help with cell culture research needs.

Impact of Microbial Contamination and Misidentified Cell Cultures on Research. Presented by Yvonne Reid, PhD, Cell Biology Collection Scientist, ATCC on March 9, 2010.

Abstract: Animal cell lines are important in vitro systems and tools for scientists in diverse disciplines beyond basic cell biology. Cell line authentication and characterization are crucial in these fields, yet most research scientists under appreciate them. Over the years, numerous cell lines have been shown to be misidentified due, in part, to poor techniques, inadequate authentication protocols, and sharing of unauthenticated cell lines amongst researchers. Technological advances have given rise to improved capabilities. Cell line authentication and characterization now require a comprehensive strategy that employs several complementary technologies for systematic testing for morphology, microbial contaminations, cellular cross-contamination as well as functionality. The validity of conclusions drawn from research data is dependent on consistent and unequivocal verification of cell line identity and function. It is estimated that the financial loss incurred by poorly characterized or misidentified cell lines is in the millions of dollars. An overview of the current technologies used to authenticate and characterize animal cell lines was presented. (Hosted by Corning)