The issue of misidentified cell lines is nothing new to the scientific community, but recent attention toward addressing this persistent challenge seems to have ramped up considerably as the year came to a close. As we reflect on the progress towards rectifying the problem of irreproducibility in science and the need for standards in addressing this issue, several events during the past year indicate considerable strides in key areas:

1. Awareness: Many times a cautionary message is only taken seriously when financial risk is quantitated. In the article, “The Economics of Reproducibility in Preclinical Research” (PLos Biol 13(6):e1002165, 2015.), Freedman et al take a deep dive into the economic impact of irreproducibility in science with an estimate of $28 B annually, in the US alone, is spent on irreproducible pre-clinical research. In a looking specifically at cell-based research, Freedman and colleagues reported that while quality control of cell lines used in biomedical research is essential to ensure reproducibility, misidentification of cell lines remains a serious problem with $100 M of research funding having potentially been spent using one misidentified cell line alone (Nat Methods 12(6): 493-7, 2015.). In efforts to even further increase awareness, the Global Biological Standards Institute (GBSI) launched the #authenticate campaign to raise awareness around the importance of authenticating cells before, during, and after research.

2. Implementation: NIH was a key driver of significant progress in implementing changes in developing policies pertaining to authentication of research materials to drive reproducible science. In 2014 NIH published Principles and Guidelines for Reporting Preclinical Research including guidelines for reporting authentication of cell lines. By 2015, over 130 editors of scientific journals, associations, and societies have agreed to endorse these principles and either require or recommend cell line authentication as condition of publication. The NIH notice (NOT-OD-15-103) on Enhancing Reproducibility through Rigor and Transparency includes provisions recommending demonstration of cell line authentication from all investigators applying for Agency funding. The guidelines become

effective on January 16, 2016. As part of an ongoing implementation push, the NIH held a Workshop on Reproducibility and Cell Culture Studies was held on September 28-29, with Yvonne Reid, Ph.D., Manager/Scientist, ATCC, an active participant in that discussion.

3. ATCC SDO efforts: Through the diligent and much appreciated effort of our work group members, ATCC SDO has recently completed and published the consensus standard: Species-Level Identification of Animal Cells through Mitochondrial Cytochrome c Oxidase Subunit 1 (CO1) DNA Barcodes (ANSI/ASN-0003-2015), for interspecies cell line identity, based on sequence technology. We look forward to the wider adoption and use of this authentication tool currently in use by cell banks, core labs and others. A related standard: ASN-0004-Species-Level Identification, and Cross-Contamination Screening in Animal Cells by Multiplex PCR is now in development.

Thank you again to the ATCC SDO working group members and others in the scientific community who are committed to standards development and utilization in support of reproducible science. Starting with increased awareness of the problem, pressure from funding agencies and journal editors to authenticate research material, together with the development of effective technology and tools, cell authentication can be a model of progress in supporting reproducibility in science.

Happy New Year to all!

Maryellen de Mars, Ph.D.

Senior Director, Standards Resource Center, ATCC

Volume 3Issue 2

ATCC® Standards Development Organization Newsletter

IN THIS ISSUE:CSP Focus 2Update on ASN-0003 3International Barcode of Life Conference 3 A Few Words About Barcoding from the ASN-0004 Workgroup Chair 4 ATCC SDO Re Accredited 5IsBioTech 3rd Fall Meeting 6NIH workshop on Reproducibility in Cell Culture Studies 7Validating Cell Culture Workshop 7

GBSI Cell Line Authentication Campaign 8International Society for Cellular Therapy 2015 Meeting 8 KAVI User Workshop 9Revision and Reaffirmation of ANSI Standard 9World Standards Week 2015 10ATCC SDO Welcomes New CSP Members 11Frequently Asked Questions 12

Ray Nims, Ph.D.Senior ConsultantRMC Pharmaceutical Solutions, Inc.Suite A, 1851 Lefthand CircleLongmont, CO 80501

Please describe your professional background, education, and how you became involved with standards.

I worked in cancer research for a number of years after receiving a B.S. degree in Biology from a small college in central Minnesota. During this time, I did lab animal (rats!) work, cell culture, and biochemical assays. I loved this time in my career, but forces beyond my control eventually pushed me back to graduate school at American University. There, I studied toxicology and pharmacology while continuing to work in the area of cancer research - now specializing in pharmacokinetics and drug metabolism. Once I had attained a Ph.D. in Chemistry, I moved into the contract testing realm, first in the field of in vitro toxicology and then in a totally new direction, lot release testing (cell line authentication and viral safety) for biopharmaceuticals. These soon became my favorite subjects.

All of this confirmed what I was told years back: “don’t worry about what subject area to get your Ph.D. in, just get it!” My technical background and photography experience came into play when I was told to develop the isoenzyme analysis assay for use in cell line identity testing at BioReliance. Photography experience, you say? Yes, staining electrophoresis gels turned out to be just like developing black and white pictures. In either case, you stop the development when the image is just right!

This is a long-winded way of explaining how I became involved in Standards. After years of testing the identity of customer cell lines using this assay, I ran into Professor Roland Nardone and was enlisted into his personal “cause” of fixing the problem of cell line misidentification. One thing led to another, and here I am

years later pitching in to help out with ASN-0002 (STR profiling), ASN-0003 (cell line barcoding), and now ASN-0004 (multiplex PCR for animal cell cross-contamination). My involvement in International Cell Line Authentication Committee (ICLAC) also grew out of the association with ATCC Standards Development Organization (SDO).

Can you describe the type of work you do in your current position?

As a consultant, I am tasked to do jobs that my pharmaceutical/biopharmaceutical clients do not have time or desire to do themselves. Long reports come to mind, who has time for that? Sometimes I am asked to help out with contamination investigations (microbial or cell line misidentification). This is the fun stuff, but like the Maytag repairman, you might have to wait a long time for such a job. So it amounts to a lot of report writing, number crunching, and the occasional white paper. I love the variety.

How did you first become involved with the ATCC SDO?

The cell line misidentification world has, until recently, involved a fairly limited group of individuals. We all know each other and we know who is willing to roll up their sleeves and work. I guess that is the best answer to this question. But seriously, I have felt fortunate and honored to work alongside this group of dedicated individuals. Roland (Nardone)’s personal cause has now been taken up by a variety of folks from around the world, and I can honestly say that progress is finally being made.

What do you see as the emerging trends or topics that will require standardized methods in the future?

I have argued that identity testing is just the tip of the iceberg when it comes to cell culture. We started there because the problem of misidentification was so rampant and costly in terms of wasted effort. But there are a variety of other

topics that could use standardization, and we are beginning to think about these now. One such area is the practice of gamma irradiation of animal serum for mitigating adventitious agent risk (e.g., viruses and mycoplasma). This is somewhat of a black art and we are in the beginning stages of identifying best practices and areas of potential standardization. Other areas that beg for standardization are the determination of population doubling time for cells and the determination of ploidy (the number of sets of chromosome pairs for the cell line).

Anything you wish to add?

I encourage subject matter experts to become involved in the standard-setting process. The committee work and involvement with an international group of like-minded individuals can be very rewarding (professionally, even if not financially).

CSP FocusCSP Focus highlights ATCC® Standards Development Organization members from different areas of the life science community and describes the contributions they are making to standards development through the ATCC SDO and their daily work.

Dr. Ray Nims

Page 2

The ATCC® Standards Development Organization recently received ANSI approval of its third consensus standard titled ASN-0003, Species-Level Identification of Animal Cells through Mitochondrial Cytochrome c Oxidase Subunit 1 (CO1) DNA Barcodes on Wednesday December 22, 2015.

The workgroup, under the leadership of Jason Katz Cooper and Yvonne Reid, Ph.D., worked tirelessly to accomplish this goal. This standard represents a groundbreaking accomplishment by the working group and the SDO, by providing the scientific community a DNA sequence-based standard that can be used for the identification of vertebrate animal cells at the species level.

To order the ASN-0003 standard, visit the ANSI eStandards Store at http://webstore.ansi.org

ASN-0003, Species-Level Identification of Animal Cells through Mitochondrial Cytochrome c Oxidase Subunit 1 (CO1) DNA Barcodes Gets ANSI Approved

International Barcode of Life Conference Authors: Mark Stoeckle and Robert HannerStarting from workshops held at Cold Spring Harbor in 2003 DNA barcoding, which examines all eukaryotic kingdoms, the conference has grown into one of the largest international research programs in biodiversity science. In August 2015, the 6th International Barcode of Life Conference, with 600 delegates from 50 countries, was held at the University of Guelph in Ontario, Canada. The conference featured 258 oral and 158 poster presentations, preconference workshops on next generation sequencing, Barcode of Life Data Systems (BOLD), formation of an International Society for DNA barcoding, and the chance to participate in a DNA barcode-based bioblitz in a nearby urban park.

In addition to the scientific program, the conference celebrated the many contributions of University of Guelph’s Dr. Paul Hebert, who first proposed DNA barcoding as a standardized DNA-based approach to species identification in 2003 and has been a vital force in organizing the initiative ever since.

The title of the 6th Conference, Barcodes to Biomes, signaled the ongoing expansion of the community’s research agenda from studies on particular sets of species in particular places to analyzing entire biotic assemblies at local and global scales. The scientific program included 30 Keynote and Plenary presentations delivered by an international array of celebrated biodiversity researchers. A common research theme was how routine DNA sequencing, in concert with a well-established DNA barcode library, enables investigating previously intractable ecological and evolutionary questions. On the applications side, the conference highlighted the increasing role of DNA barcoding in food authentication and regulatory interests, ranging from trade in endangered species to herbal supplements.

Dr. Paul Hebert proposed extending the international collaborative efforts toward a grand “Planetary Biodiversity Mission”, with the

aim of expanding the DNA barcode library from 500 thousand to 10-20 million species over the next two decades.

The pre-conference bioblitz at the rare Charitable Trust Reserve, a 900 acre preserve in nearby Cambridge, Ontario, yielded a published article by the end of the conference (www.dnabarcodes2015.org/bioblitz/). A comprehensive sampling scheme over four months, followed by the one-day bioblitz, resulted in 25,287 specimens barcoded and 3,502 samples vouchered at the Biodiversity Institute of Ontario collection, covering 14 phyla, 29 classes, 117 orders, and 531 families of animals, plants, fungi, and lichens.

For more information, the conference program, abstracts, and streaming videos of Plenary Sessions are available online (www.dnabarcodes2015.org).

Page 3

How can one be certain of the identity of a cell growing in culture? Two cells of the same type derived from different specimens can be impossible to distinguish through a microscope. This is the case whether the cells are conspecific or whether they are from entirely different species. A cell in vitro may or may not have the same identity indicated on the vial label. After paying a great price in lost time and effort from misidentified cell lines, the research community now identifies human cells by genetic analysis of short tandem repeats (STR). STR analysis is accurate, relatively fast and inexpensive, and ensures the integrity of human cell culture systems. However, STR does not identify species origin. This can be a big issue particularly in labs using multiple cell lines for research or bioproduction.

To address this concern, a new standard working group, ASN-0004 Species-Level Identification and Cross-Contamination Screening in Animal Cells by Multiplex PCR, is being launched. Our goal is to develop a standard method that will complement STR analysis. To this end, our strategy is to employ primer pairs that specifically recognize one species. To accomplish this we suggest targeting mitochondrial genes that tend to be highly conserved within a species but diverge between species. With this strategy a simple multiplex PCR reaction can be standardized to provide basic species identification and safeguard against cross contamination. When combined with STR profiling, scientists of the near future, unlike those of the recent past, will read the labels on cell culture vials with supreme confidence in the identity of the contents.

About Jason Cooper

Mr. Cooper is currently a Biology Professor (Adjunct) in Pittsburgh, PA and has worked as a scientist, researcher, and supervisor at Roche Molecular Systems, Aptagen, Inc., and the American Type Culture Collection (ATCC). His publications, presentation, and work include:

• Kher CP, Doeder P, Cooper J, Ikonomi P, Achilles-Day U, Kupper F, Lynn H. Barcoding Tetrahymena: discriminating species and identifying unknowns using the cytochrome c oxidase subunit 1 (cox-1) barcode. Protist 162(1):2-13, 2011.

• Cooper JK, Cottrill K, Ivanova NV, Hanner R, Ikonomi P. A Two-pronged molecular approach to species identification and detection of cross-contamination in human and animal cell lines. In Vitro Cell Dev Biol Anim 43(10):344-351, 2007.

• Fernandes AS, Le T, Cooper JK, Ault M, Otaizo-Carrasquero F, Irish T. Producing high quality genomic DNA from a wide variety of microbes. American Society for Microbiology (ASM) annual meeting, 2008.

• Cooper J, Ivanova NV, Ikonomi P, Hanner R. DNA Barcodes for definitive species identification of cells in culture. Amercian Society of Cell Biology (ASCB) annual meeting, 2006.

• Cooper J, King S, Cottrill K, Sykes G, Crissup M, Ikonomi P. Detection of interspecies cross-contamination in cell culture: A highly sensitive multiplex PCR based approach. American Society of Cell Biology (ASCB) annual meeting, 2006.

Page 4

Workshops & Presentations

A Few Words About Barcoding from the ASN-0004 Workgroup Chair

Page 4

Workshops & Presentations

(continued)

ATCC SDO Undergoes Reaccreditation ATCC has been an American National Standards Institute (ANSI)-accredited standards developer since 2007, and was the first and only Biological Resource Center (BRC) to become an ANSI-accredited Standards Development Organization (SDO). ANSI is an internationally recognized organization that accredits standards development organizations. The not-for-profit ANSI has been designated by the U.S. government as the administrator of voluntary consensus standards, known in this country as American National Standards. ATCC® published standards are recognized by ANSI and are compatible with requirements of the International Organization for Standardization. SDO status enables ATCC to lead development of best practices for use in the life science industry and to promote their use globally, using a consensus driven process that balances the viewpoints of industry, government, regulatory organizations, and academia.

ANSI reaccreditation is required yearly to ensure that the policies and procedures of the ATCC SDO are aligned with the ANSI Essential Requirements. This reaccreditation also includes periodic audits by ANSI to thoroughly review and follow the SDO process.

This past summer, the ATCC SDO underwent such an audit conducted by ANSI auditors Jay Moskowitz, Director, ANSI Standards Developer Audit Program, and Mickie Rops from Mickie Rops Consulting LLC. This provided an opportunity for a review of the operations of ATCC as related to standards development and associated activities, including continuity of administrative oversight and support of the standards activities. Questions and answers from both parties, as well as the discussion of any findings, recommendations, and comments were addressed during a post audit teleconference call. Final recommendations were presented to the ANSI Executive Standard Council’s Audit Subcommittee on January 14, 2016, and the ATC SDO is awaiting the decision.

Page 5

IsBioTech 3rd Fall Meeting The IsBioTech 3rd Fall Meeting on Cell Banking, Contaminant Control, and Process Analysis and Automation, was held on September 28-30, 2015, in Rosslyn, Virginia.

Dedicated scientists and engineers who have the daily responsibilities of creating and managing procedures and assays for reproducible outcomes were invited to attend. Through these presentations, highly innovative techniques/technologies and best practices relevant to creating well-characterized cell banks, preventing and identifying contaminants during the process and testing methods for rapid scale-up of clinical materials and licensed products were shared.

This year’s program chair for the Cell Banking sessions was Dr. Yvonne Reid, Manager, Scientist ATCC. Some of the featured presentations and speakers included

• Haruhiko Murata, M.D., Ph.D. Research Biologist, Food and Drug Administration Center for Biologics Evaluation and Research Qualification of Madin-Darby Canine Kidney Cells for Flu Vaccines

• Linda Fröderberg-Roth, Ph.D. Senior Research Scientist, AstraZeneca R&D Cell Banking for Pharmaceutical Research

• Jamie Almeida Research Biologist, Biochemical Science Division, Bioassay Methods Group National Institute of Standards & Technology (NIST) NIST’s Efforts in Non-Human Cell Line Authentication

• Gregory Sykes Lead Biologist, QC Testing, ATCC Mitochondrial Multiplex and COI Barcode Analyses as Alternatives to Isoenzymology

• Afshin Sohrabi, Ph.D. Senior Scientist, Molecular Development Services, BioReliance Corporation Molecular Characterization of Cell Lines

• John M. Baust, Ph.D. President and Lead Scientist, CPSI Biotech Continued Development of Novel Freezing and Thawing Devices for Improving Cell Cryopreservation

Workshops & Presentations

Dr. Yvonne Reid, ATCC

Page 6

On October 2015, approximately 80 scientists participated in a one-day workshop in London, England focused on validating cell culture. The workshop organised jointly by the European Collection of Authenticated Cell Cultures (ECACC) and Sigma-Aldrich, and run by ECACC.

The workshop presentations focused on the importance of good cell culture practice and authentication testing for experimental work and future funding. Choosing the right cell line, ensuring the cell line is fit for purpose, and understanding how culture conditions can influence function and physiology, were all explored in detail.

The workshop brought together a group of scientists with expertise in cell culture and that were clearly passionate about cell culture authentication and validation. Members of the International Cell Line Authentication Committee (ICLAC), including Ed Burnett, Amanda Capes-Davis, Jim Cooper, and Lyn Healy, enjoyed the chance to speak and participate in discussing topics such as,

how to test cultures for mycoplasma and how often to test for contamination.

The US National Institute of General Medical Sciences (NIGMS) under the leadership of Dr. Jon Lorsch, and Dr. James Deatherage, recently hosted a NIH workshop on Reproducibility in Cell Culture Studies on September 28-29, 2015. At this meeting, experts from academia, industry, funding agencies, and journals came together to discuss how to address issues with reproducibility in studies that utilize cell lines. In addition, the workshop attendees debated how to identify current quality control challenges and new opportunities in cell-based research. The workshop also included panel discussions on cell line identification, genetic and phenotypic characterization of cells, phenotypic and genotypic heterogeneity of cells, reagents, and best practices in research and reporting.

A general consensus was reached that a community-wide change in cell culture practices is needed, which could be realized by training and awareness. It was also recognized that solutions to many of the issues already exist and that to stimulate meaningful change requires the key stakeholders (i.e., journals, funding agencies, academia) to convene and agree to enforce policies of best practices and thorough reporting. The workshop concluded by composing key recommendations designed to guide stakeholders in this decision making process. A manuscript outlining these recommendations is in preparation.

Some of the session chairs included, Dr. Richard Neve, Senior Research Scientist, Gilead Sciences, Inc., Dr. Roland Nardone, Professor Emeritus, Catholic University of America, and Dr. Yvonne Reid, Manager Scientist, ATCC.

For more information, visit: loop.nigms.nih.gov/2015/09/nih-workshop-on-reproducibility-in-cell-culture-studies/.

NIH workshop on Reproducibility in Cell Culture Studies

Workshops & Presentations

(continued)

Dr. Richard Neve, Senior Research Scientist, Gilead Sciences, Inc.

Dr. Roland Nardone, Professor Emeritus, Catholic Univ. of America

Validating Cell Culture Workshop Authors: Amanda Capes-Davis and Lyn Healy

Speakers at the Validating Cell Culture Workshop (from left): Bryan Bolton, Jim Cooper, Amanda Capes-Davis, Valerie Speirs, Jamie Meredith, Lyn Healy, Ed Burnett. Source: Twitter @ECACC.

Page 7

GBSI Cell Line Authentication Campaign GBSI’s #authenticate awareness campaign continues to build awareness and advance policies that will make cell line authentication a routine and integral part of good research practice. To date, the campaign has engaged researchers, academics, private and government funders, journals, scientific societies, cell authentication service providers, and other stakeholders, as well as multiple champions and sponsors—including ATCC. Throughout 2015, the number and caliber of scientific and media articles on the importance of authentication has been tremendous. The campaign is making a difference as we see NIH and other funders incorporating cell authentication into their grant application and review guidelines, while Nature and other biomedical journals continue strengthening their policies on cell authentication. But until cell line authentication becomes a routine and documented part of good research practices for cell culture, our work is not done.

The campaign’s Cell Culture and Authentication Survey results were recently published in BioTechniques and received extensive media coverage. A few of the key results:

• Of the 446 total survey respondents, 52% said that they did not authenticate their cells, citing barriers to performing cell line authentication as cost (61%), time (53%), and delays in research (35%)

• Most respondents noted the need for additional training; only 62% received training on the problems associated with cell line misidentification

• Almost 75% of the survey respondents support development and use of additional standardized techniques for authenticating cell lines

In addition to publishing and sharing the survey results, GBSI continues to advance the conversation. Work is underway to develop our cell authentication training module funded by NIH with generous support from Susan G. Komen. The campaign raised the bar on this topic and is evolving into a soon to launch Cell Authentication Alliance, which will bring together a broader coalition of cell line authentication stakeholders in a virtual, efficient, and active network.

International Society for Cellular Therapy 2015 Meeting The International Society for Cellular Therapy (ISCT) held its 21st Annual Meeting at Caesars Palace in Las Vegas, Nevada the week of May 27, 2015.

Clinicians, regulators, technologists, and industry partners with a shared vision to translate cellular therapy into safe and effective therapies to improve patients’ lives attended this meeting. Dr. Leonard Freedman, President of the Global Biological Standards Institute, opened the workshop with a presentation about the case for standards in life science research. Subsequent sessions included: Presentations about instrumentation and reagents; standardization of data analysis; commercial product development; and flow-based, in-process and release testing by representatives from academia, regulatory organizations, clinical institutes, and industry. The workshop sparked a renewed focus and dialogue on standardization in flow cytometry that will continue at the next flow cytometry workshop during the 2016 ISCT annual meeting in Singapore.

For more information on this meeting:

http://www.celltherapysociety.org/event/id/494270/21st-ISCT-Annual-Meeting.htm

Workshops & Presentations (continued)

Page 8

On Tuesday, December 8, 2015, Kavi Corporation hosted its Annual Kavi® User Group workshop at the American Society of Clinical Oncology Conference Center in Alexandria, VA. The user group workshop provided an opportunity to share the company’s vision, product progress, and development plans that would assist the SDO users. The interactive setting also allowed users to share information and tips on how to better serve the SDO community and get workgroup members to feel comfortable and utilize the member workspace for standards development.

Kavi Workspace is a unique, integrated, easy-to-use solution that helps manage the standards process, increase engagement, and achieve consensus faster.

Revision and Reaffirmation of ANSI Standard In late July 2015, the workgroup for ASN-0001 reconvened to update the SDO’s first ANSI-accepted standard titled ASN-0001, Standardization of in vitro Assays to Determine Anthrax Toxin Activities.

This new updated version of the standard was again led by Molly A. Hughes, M.D., Ph.D., Associate Professor of Medicine, Division of Infectious Diseases, at the University of Virginia School of Medicine. This revision represents the second version of ASN-0001, reflecting changes and updates in the five-year period since the first ASN was published. This standard provides a reference method for performing in vitro assays to determine anthrax toxin activities. The anthrax toxins to be assayed are lethal toxin [LT; protective antigen (PA) + lethal factor (LF)] and edema toxin [ET; PA + edema factor (EF)]. The assay for LT is based on cytotoxicity to cultured murine macrophages. The assay for ET is based on its adenylate cyclase activity and cyclic adenosine monophosphate production in host cells. These assays may be used to determine activities of various LT and ET preparations for experimental comparison between the same or different groups of investigators.

Under the new designation, ATCC ASN-0001.1, Standardization of in vitro Assays to Determine Anthrax Toxin Activities (revision and redesignation of ANSI/ATCC ASN-0001-2009), this standard represents a ground breaking accomplishment by the working group and the SDO.

To purchase this and any published SDO standard, please visit webstore.ansi.org.

KAVI User Workshop

Page 9

World Standards Week 2015The ATCC SDO joined the global standardization community in celebrating World Standards Week, which began on September 28 and ran through October 2, 2015. Meetings and related activities were held in various locations in the Washington DC area. World Standards Week is held annually, and pays tribute to the thousands of members around the world who participate in standardization activities and help to raise awareness of the role that standards play in addressing national and global priorities. Leaders from the standardization community were in attendance throughout the week-long series of events that included an ANSI awards banquet and ceremony.You can find a full list of the events on http://www.ansi.org/meetings_events/wsw15/wsw.aspx?menuid=8

Page 10

ATCC SDO Welcomes

New CSP Members

Dr. Hitomi AsaharaUniversity of California, Berkeley

Sonia HoughtonAstraZeneca

Angie HutzelDNA Center

Jun XiAnhui Hefei University

Tsuyoshi KarasawaSUMMIT Pharmaceuticals International Corporation

Bernie McKayUlster University

Dr. Varsha MeghnaniCastle Medical, LLC

Dr. Richard NeveGilead Sciences, Inc

Dr. David RozakUSAMRID

Dr. Mark StoeckleRockefeller University

Angela WeberZoetis

Lekshmi Bhuvanendran Pillai Sreelatha The College of Pharmaceutical Sciences

Social MediaThe ATCC® Standards Development Organization (SDO) participates in two social media websites, Facebook and LinkedIn. The websites provide timely information regarding events and initiatives as well as provide a place for discussion about current topics, news, and articles of interest to the life science community.

Find us on. . . The ATCC SDO Facebook page includes an overview of the Standards Development Organization, upcoming events, photographs, and discussions. Become a fan of our ATCC® SDO Facebook page! Accessible to registered users of LinkedIn, the group page provides a way for members to network and share articles, upcoming workshops, websites, and information of interest.

Join our LinkedIn network!

Page 11

This newsletter is published by ATCC and is distributed free of charge upon request. Direct all correspondence to P.O. Box 1549, Manassas, VA, 20108 or e-mail standards@atcc.org. Photocopies may be made for personal or internal use without charge. This consent does not extend to copying for general distribution, promotion, creating new works, or resale.

© 2015 American Type Culture Collection. The ATCC trademark and trade name are owned by the American Type Culture Collection.

About CO1 DNA Barcode to Animal Species Identification1. Question: What is Multiplex PCR? Cytochrome c oxidase subunit I (CO1)?Answer: A multiplex PCR is a single reaction that can simultaneously amplify several targets. The power of the assay is speed and ease of use, as it can screen for the presence or absence of several targets in a mixture. CO1 is a mitochondrial gene that is highly conserved within a species but varies greatly between species. Thus, CO1 is an ideal candidate against which to engineer a multiplex screening assay with primers that are species-specific.

2. Question: How does this tie in with CO1 and STR? Answer: STR profiling and CO1 barcoding are designed to confirm the identity of a cell to the individual (STR profile) or species (CO1 barcoding) level. Neither of these assays is really designed to detect an inter-species cross-contaminating cell type. Multiplex PCR using CO1 primer sets will address this need.

3. Question: How have we detected inter-species cross-contamination in the past?Answer: Karyotyping and isoenzyme analysis were the primary methods used in the past. The disadvantages of these methods are that karyotyping is too expensive and time consuming for routine use, and the reagents needed to perform isoenzyme analysis are no longer commercially available.

4. Question: Will the Multiplex CO1 assay be able to detect any species of contaminating cell?Answer: No, this assay has been designed to detect the ten or more most commonly employed species of cells. Despite this limitation, the assay will be a very useful tool for monitoring cell lines for freedom from cross-contaminants.

5. Question: Who needs to employ this tool?Answer: Any laboratory simultaneously studying cell lines derived from a number of different animal species.

6. Question: Where can I learn more?Answer: Cooper JK, Cottrill K, Ivanova NV, Hanner R, Ikonomi P. A Two-pronged molecular approach to species identification and detection of cross-contamination in human and animal cell lines. In Vitro Cell Dev Biol Anim 43(10):344-51, 2007.

Frequently Asked Questions

Page 12