happy new year standards community!€¦ · happy new year standards community! throughout 2016,...

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Happy New Year Standards Community! Throughout 2016, ATCC continued to focus on the development and provision of standards through the ATCC Standards Development Or- ganization (SDO) and the products and services we offer. The success of the ATCC SDO relies on the devoted chairs, co-chairs, working groups, and members of the Consensus Standards Partnership (CSP) who keep efforts directed toward timely and relevant scientific issues. Thank you to the extended ATCC SDO team for the continued interest, involve- ment, and results. Key highlights from SDO activities and related ATCC work from the past year include: Mouse STR CRADA and Consortium ATCC is pleased to collaborate with the National Institute of Standards and Technology (NIST) through a three-year coop- erative research and development agreement (CRADA) to de- termine and validate short tandem repeat (STR) markers for mouse cell line authentication. Under the agreement, ATCC partnered with NIST to establish a Mouse Cell Line Authentica- tion Consortium, and will provide DNA from at least 50 mouse cell lines. ese DNA samples will be part of a kit that will be distributed to Consortium members for inter-laboratory valida- tion studies. While validated methods for human cell line iden- tification are available to the scientific community, this effort is aimed at developing similar technology for nonhuman cell lines. ASN-0002 Authentication of Human Cell Lines: Standardization of STR Profiling is pivotal consensus standard, developed through the ATCC SDO, is reaching its five-year milestone and is up for revision/ renewal. is document elaborates a standardized procedure for authentication and identification of human cell lines using STR profiling and a searchable public database. ASN-0004 Species-Level Identification and Cross-Contamination Screening in Animal Cells by Multiplex PCR Working Group is group is hard at work to develop a procedure for species- level identification and cross contamination screen¬ing for 15 species considered the most relevant in cell culture. ese spe- cies include human, mouse, Chinese and Syrian hamsters, dog, rat, pig, African Green and Rhesus monkeys, bovine, cat, horse, rabbit, chicken, and Guinea pig. A major driver for the develop- ment of this standard is the reagents for the traditional isoen- zymological identification method are no longer commercially available, and there is an urgency to fill the technology void. ATCC Representation at the Association for Molecular Pathology (AMP) Conference In November, we had the honor of presenting ATCC in Char- lotte, North Carolina at the AMP conference. Liz Kerrigan, di- rector of Commercial Partnerships and Standards, participated in the AMP Reference Material Forum highlighting ATCC’s ex- panding portfolio of Zika virus reference materials. Dr. Maryel- len de Mars, senior director of ATCC’s Standards Resource Cen- ter, was a featured speaker at the AMP International Showcase on advancing standards for clinical diagnostics and the need for global reference materials and standardization. I would like to close by saying thank you and goodbye to Dr. Yvonne Reid (featured in this issue), who retired at the end of 2016 after 35 years of devoted service to ATCC. Yvonne has been essential to the success of ATCC, where her expertise and particular dedication to cell line authentication and characterization has forever shaped cell culture quality standards. Yvonne will continue on with ATCC in a consultant capacity as she finishes up some key cell culture projects in early 2017. I wish her health and happiness in her next adventure! Maryellen de Mars, Sr. Director, Standards Resource Center Volume 4 Issue 1 ATCC ® STANDARDS DEVELOPMENT ORGANIZATION NEWSLETTER In This Issue Yvonne Reid, PhD Receives Highest Honor from The Society of In Vitro Biology ......................................................................................... 2 Eugene Elmore, PhD SIVB Lifetime Achievement Award Recipient .. 2 ASN-0004 Species-Level Identification and Cross-Contamination Screening in Animal Cells by Multiplex PCR: An Update ................... 4 The International Society for BioProcess Technology Meeting ............ 8 AMP Meeting Focuses on the Need for Standardization in the Global Market .................................................................................................... 8 KAVI User Workshop ............................................................................ 9 ANSI/ATCC ASN-0002 Revision and Reaffirmation ............................ 9 World Standards Week 2016 ................................................................ 10

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Page 1: Happy New Year Standards Community!€¦ · Happy New Year Standards Community! Throughout 2016, ATCC continued to focus on the development and provision of standards through the

Happy New Year Standards Community! Throughout 2016, ATCC continued to focus on the development and provision of standards through the ATCC Standards Development Or-ganization (SDO) and the products and services we offer. The success of the ATCC SDO relies on the devoted chairs, co-chairs, working groups, and members of the Consensus Standards Partnership (CSP) who keep efforts directed toward timely and relevant scientific issues. Thank you to the extended ATCC SDO team for the continued interest, involve-ment, and results.

Key highlights from SDO activities and related ATCC work from the past year include:

▪ Mouse STR CRADA and Consortium ATCC is pleased to collaborate with the National Institute of Standards and Technology (NIST) through a three-year coop-erative research and development agreement (CRADA) to de-termine and validate short tandem repeat (STR) markers for mouse cell line authentication. Under the agreement, ATCC partnered with NIST to establish a Mouse Cell Line Authentica-tion Consortium, and will provide DNA from at least 50 mouse cell lines. These DNA samples will be part of a kit that will be distributed to Consortium members for inter-laboratory valida-tion studies. While validated methods for human cell line iden-tification are available to the scientific community, this effort is aimed at developing similar technology for nonhuman cell lines.

▪ ASN-0002 Authentication of Human Cell Lines: Standardization of STR Profiling This pivotal consensus standard, developed through the ATCC SDO, is reaching its five-year milestone and is up for revision/renewal. This document elaborates a standardized procedure for authentication and identification of human cell lines using STR profiling and a searchable public database.

▪ ASN-0004 Species-Level Identification and Cross-Contamination Screening in Animal Cells by Multiplex PCR Working GroupThis group is hard at work to develop a procedure for species-level identification and cross contamination screen¬ing for 15 species considered the most relevant in cell culture. These spe-cies include human, mouse, Chinese and Syrian hamsters, dog, rat, pig, African Green and Rhesus monkeys, bovine, cat, horse, rabbit, chicken, and Guinea pig. A major driver for the develop-ment of this standard is the reagents for the traditional isoen-zymological identification method are no longer commercially available, and there is an urgency to fill the technology void.

▪ ATCC Representation at the Association for Molecular Pathology (AMP) Conference In November, we had the honor of presenting ATCC in Char-lotte, North Carolina at the AMP conference. Liz Kerrigan, di-rector of Commercial Partnerships and Standards, participated in the AMP Reference Material Forum highlighting ATCC’s ex-panding portfolio of Zika virus reference materials. Dr. Maryel-len de Mars, senior director of ATCC’s Standards Resource Cen-ter, was a featured speaker at the AMP International Showcase on advancing standards for clinical diagnostics and the need for global reference materials and standardization.

I would like to close by saying thank you and goodbye to Dr. Yvonne Reid (featured in this issue), who retired at the end of 2016 after 35 years of devoted service to ATCC. Yvonne has been essential to the success of ATCC, where her expertise and particular dedication to cell line authentication and characterization has forever shaped cell culture quality standards. Yvonne will continue on with ATCC in a consultant capacity as she finishes up some key cell culture projects in early 2017. I wish her health and happiness in her next adventure!

Maryellen de Mars,Sr. Director, Standards Resource Center

Volume 4Issue 1

ATCC® STAndArdS developmenT orgAnizATion newSleTTer

In This IssueYvonne Reid, PhD Receives Highest Honor from The Society of In Vitro Biology ......................................................................................... 2

Eugene Elmore, PhD SIVB Lifetime Achievement Award Recipient .. 2

ASN-0004 Species-Level Identification and Cross-Contamination Screening in Animal Cells by Multiplex PCR: An Update ................... 4

The International Society for BioProcess Technology Meeting ............ 8

AMP Meeting Focuses on the Need for Standardization in the Global Market .................................................................................................... 8

KAVI User Workshop ............................................................................ 9

ANSI/ATCC ASN-0002 Revision and Reaffirmation ............................ 9

World Standards Week 2016 ................................................................ 10

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Yvonne Reid, PhD Receives Highest Honor from The Society of In Vitro Biology

The 2016 Distinguished Lifetime Achievement Award from the Society for In Vitro Biology (SIVB) was awarded to Dr. Yvonne Reid for her pioneering work in short tandem repeat (STR) DNA profiling for human cell line identification. This award is the highest honor given by SIVB, and is presented annually to a scientist who has made significant contributions to the field of biology.

Dr. Reid has over 30 years of expertise in the field of cell biology, immunology, and molecular biology. She has authored more than 41 peer-reviewed publications, served on countless sci-entific committees, and has received numerous awards throughout her very impressive career.

Dr. Reid’s impeccable reputation as a cell culture expert and fierce proponent of cell line authen-tication and characterization has won her both national and international acclaim. She has been a mentor to countless cell culturists, emphasizing the importance of good lab practices. Dr. Reid has been crucial to the success of the American Type Culture Collection (ATCC), where she has worked as a manager and collection scientist in the Cell Biology program for 35 years. Her contributions and forward thinking has propelled her to testify before the subcommittee on Inves-tigations and Oversight of the U.S. House of Representatives to discuss cell line authentication.

Dr. Reid is a herald champion and crusader, and we salute her for her significant scientific achievements in the development of cell culture and science!

Eugene Elmore, PhD SIVB Lifetime Achievement Award Recipient The other SIVB Lifetime Achievement Award was presented to Dr. Eugene Elmore. Dr. Elmore retired as a project scientist from the Department of Radiation Oncology at the University of California Irvine, in Irvine, Ca. His career includes serving as a co-organizer of the US EPA Conference on Mechanisms of Gap Junction Function and Relevance to Disease. In addition, he was a vital contributor and workgroup member to the ANSI standard ASN-0002 titled Authen-tication of Human Cell Lines: Standardization of STR Profiling. From 2010 to 2012 Dr. Elmore served on the Scientific Advisory Committee on Alternative Toxicological Methods (SACATM), ICCVAM/NICEATM, NTP, NIEHS. Dr. Elmore’s contributions to the scientific community also includes his duties as the scientific expert for the European Commission’s European Centre for the Validation of Alternative Methods (ECVAM) Science Advisory Committee and Workgroup in Ispra, Italy, as well as numerous awards and accolades by the SIVB that include his receiving the 2004 Fellow Award recipient for the SIVB, the 2013 Distinguished Service Award, and his service as the SIVB President Elect for the 2012-2014 term.

SIVB established the Lifetime Achievement Award in 1989 in order to recognize the outstand-ing contributions from scientists who have made significant contributions to the field of in vitro biology and/or in the development of novel technologies that have advanced in vitro biology. Dr. Elmore is truly a trailblazer in the scientific field!

Awards

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Feature SDO Member Yvonne Reid, PhD

Yvonne Reid, PhDManager, Scientist, ATCC10801 University BlvdManassas, VA 20110

Please describe your professional back-ground, education, and how you became involved with standards. I received my PhD in Zoology from Howard University, Washington DC, in 1985, five years after joining ATCC. During my first few years at ATCC, with a background in immunology, my research focus was on the development and characterization of several monoclonal an-tibodies and their epitopes that targeted endo-thelial and epithelial-associated antigens. Shortly after I received my PhD, I was as-signed a project to apply DNA fingerprint-ing technology to the ATCC general collec-tion of human cell lines using the multi-locus probe, 33.6, developed by Sir Alec Jeffreys. This new technology was recommended by Dr. Steve O’Brien, who served as Chief of the Laboratory of Genomic Diversity at the National Cancer Institute (NCI), National In-stitutes of Health (NIH) from 1986 to 2011, as well as a member of the Cell Biology Ad-visory Committee at the ATCC. I commuted to Dr. O’Brien’s laboratory for several weeks to learn the DNA fingerprinting technique, which we soon applied to our cell bank. We found that the complex banding patterns of the multi-locus probes, although informative, were not reproducible. We then applied Vari-able number tandem repeat (VNTR) markers. The VNTR markers were less informative than the multi-locus probes, the procedure was time consuming, and lacked sensitivity; however, it provided discreet bands that were easily interpreted. By the mid-1990s, we had access to the STR markers which were eas-ily adaptable to PCR. We started with 4 STR markers, then expanded to 7 STR markers + amelogenin, and the currently used procedure includes 17 STR markers + amelogenin. In the early years of the project, I reached out to experts in the forensic community by visiting their laboratories, attending scientific meet-ings, sharing data, and absorbing their input

and suggestions. Subsequently, I became the collection scientist for the ATCC cell biology program. I was re-sponsible for the acquisition, expansion, char-acterization, and distribution of all animal cell lines and hybridomas. It was during this time that I recognized that some of the existing hu-man cell lines and newly acquired human cell lines were misidentified. Compared to previ-ous technologies such as isoenzymology and cytogenetic analysis (i.e., karyotyping, whole chromosome paint, SKY), the new technolo-gies (i.e., DNA fingerprinting, VNTR and STR analysis) were more informative in identify-ing each human cell line. This consequently revealed misidentified cell lines within the collection, most of which were deaccessioned. Research in the cell biology program also led to the application of the CO1 barcoding for interspecies identification of animal cell lines and PCR analysis for mycoplasma detection.

Despite the fact that by the mid-90s we had better reagents, supplies, equipment and tech-niques, misidentification of newly acquired human cell lines persisted. I began to spread the word through publications, workshops, webinars, and platform presentations at sci-entific societies, and promote the use of best practices in cell culture in order to reduce the misidentification of human cell lines, thus en-suring reproducible results. The application of STR markers at ATCC in the mid-1990s and subsequently at other cell banks and laboratories in the scientific community, has led to the development of a written consensus standard (ASN-0002) on the use of STR profil-ing for human cell line identification.

Can you describe the type of work you do in your current position?As Manager/Scientist of the Standard Re-source Center (SRC), I am currently involved in several initiatives pertaining to standard development in cell banking, focusing on best practices in cell culture, and the authentica-tion of human and non-human cell lines – all of which will impact reproducible results.

How did you first become involved with the ATCC SDO?I first became involved with the ATCC SDO

when I was nominated by the members of a working group tasked to develop a consen-sus standard approved and accepted by the American National Standard Institute (ANSI) on cell line authentication. I was asked to serve as the work group co-chair. The authen-tication method selected was STR, and the standard that was eventually developed and accepted by ANSI is titled ASN-0002- Au-thentication of Human Cell Lines: Standard-ization of STR Profiling. This international working group of experts had a tremendous amount of experience in forensic science and STR profiling of human cell lines, and made the entire standard development encounter in-valuable. The working group had representa-tion from academia, government, regulatory, industry, and from cell banks.

What do you see as the emerging trends or topics that will require standardized methods in the future?New data has emerged suggesting that use of 7 STR markers + amelogenin is not sufficient to discriminate among human cell lines. Un-deniably, the application of more than 17 STR markers may be necessary for unique identity of human cell lines. SNPs and Next Genera-tion Sequencing are mature technologies that will become widely used for animal cell line speciation, both interspecies and intraspecies. The consolidation of information from these technologies in a free-access public database would serve the scientific public well. These new technologies and indeed emerging tech-nologies, should be standardized and validat-ed for their robustness to confirm their claims.

Anything you wish to add? I think it is important to view cell line authen-tication as more than cell line speciation or cell line identity. One can define an authentic cell line as a cell line species that retains its unique characteristics and functionality as de-fined by its purpose of use. It is not surprising that sub-optimal growth conditions, species misidentification, and the contamination of cultures with adventitious agents, are a few of the causes that can impact cell behavior and authenticity--- all of which can lead to irre-producible results.

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ASN-0004 Species-Level Identification and Cross-Contamination Screening in Animal Cells by Multiplex PCR: An Update

Everyone is looking for a few extra hours in the day and a couple of extra dollars. Yet, when a new cell line is acquired, it is highly recommended not to cut too many corners, make sure the identity of the cell line is cor-

rect, and that the cell line is free of cross-con-taminating cells. Our goal with the current consensus standard in development, ASN-0004 Species-Level Identification and Cross-Contamination Screening in Animal Cells by Multiplex PCR, is to recommend a procedure for species-level identification and cross con-tamination screening that balances the need for testing rigor, while keeping speed and cost efficiency at the forefront.

Traditionally, isoenzymological identification methods have been useful for species identifi-cation in cell culture, but the reagents for this method are no longer commercially available. In its place, we are recommending a multiplex PCR-based approach. The approach is sim-ple and painless, only requiring the technol-ogy needed for standard PCR. Primers in the multiplex are designed to produce amplicons only when the target species is present in the culture. The resulting amplicons vary in size

as well as in a species-specific manner. This way, an analyst can quickly screen a large number of cultures and be able to identify the species by simply looking at the band size.

The work group has identified 15 species that we consider to be most relevant to the cell culture community. This includes human, mouse, Chinese and Syrian hamsters, dog, rat, pig, African Green and Rhesus monkies, bo-vine, cat, horse, rabbit, chicken, and Guinea pig. Currently, members of the work group are fine tuning and finalizing the optimal con-ditions for the assay. This step has prompted a couple work group members to go back to the lab and work. We are confident that this methodology will soon fill the void left by isoenzymology, and will become a standard practice in universities and industry, and pro-vide the research community with everything they need for the most confident results (short of extra cash and more hours in the day!).

Jason Cooper, PhD

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Utilizing the ATCC SDO Website

The ATCC SDO has a designated webpage and working webspace just for you! Browse the public pages to discover the history, for-mation, mission, and vision of the SDO, as well as listing the SDO organization chart, and the SDO Policy and Procedure manual:

As an SDO member, you can log in to the members only pages to view the SDO news and announcements, suggest new standards, and even cast your ballot for new or revised standards. As a member of the ATCC SDO, getting to the members only pages is easy…just need to log in!

On the landing page/ Home Page, go to Login located on the top right hand or you can log in from the Log in prompt on blue on the left hand side:

Once the log in page appears, enter your Log-in User name and password. In the event this is the first time logging in, you will need to request a password.

NOTE: Please contact the SDO administra-tor via e mail if you cannot remember your log in name—that information will need to be sent to you separately. In addition, make sure that your contact information is correct, email, affiliation, etc, or the system will not recognize you.

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Once logged in, the SDO Member’s Only landing page will appear, along with:

▪ Your personalized welcome in the top right hand corner of the page

▪ Your company affiliation

▪ The bulletin board with notices

▪ Your groups tab (in this list are any subgroups or workgroups that you are currently assigned to)

▪ Your project review tab (if you are currently assigned to a project)

▪ Your Take Action tab (any tasks requiring your attention right now):

▪ The SDO Policy and Procedure Manual

▪ CSP Member’s Hand book

▪ CSP membership list

▪ A training center if you are interested in knowing more

There are several ways that you can view the document and ballot to cast your vote.

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The quickest way to cast your ballot would be to go to the Take Action tab:

You can also go to the Ballots Open tab to bring it up:

Or you can go under your Workspace tab, select Groups to open up the box that shows all of the workgroups that you belong to, and select ballots:

If you need assistance, please contact [email protected]….always glad to help!

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The International Society for BioProcess Technology MeetingThe International Society for BioProcess Technology (ISBioTech) held its 4th Fall Meeting in Virginia Beach, Va from December 12-14, 2016. The meeting was designed for the professionals responsible for devel-oping and producing biologics worldwide, and allowed participants to share the techniques and best practices needed to establish stable and well-characterized cell banks. Three concurrent programs on Cell & Tissue Banking, Contaminant Control, and Process Analysis & Au-tomation were discussed for a wide-range of applications including vaccines, antibodies, recombinant proteins, viral vectors, and cellular therapies.

Some of the featured presentations included:

▪ Jamie Almeida • National Institute of Standards & Technology – Mouse Cell Line Authentication Consortium

▪ Yvonne A. Reid, Ph.D. • American Type Culture Collection (ATCC) – Comprehensive Overview of Cell Line Authentication for Pre-Clinical Research and DNA Typing of Human Cell Lines: Historical Perspective

▪ Gregory R. Sykes • American Type Culture Collection (ATCC) – Mitochondria-Targeting PCR and CO1 Barcode Sequence Analyses as Alternatives to Isoenzymology

▪ John M. Baust, Ph.D. • CPSI Biotech Improving Processing and Recovery of Cryopreserved Cell and Tissues

▪ Raymond Nims, Ph.D. • RMC Pharmaceutical Solutions, Inc. – Worst Case Model Viruses for Low and High pH Inactivation

For more information on this and other meetings sponsored by ISBioTech, please visit the ISBioTech website at http://www.isbiotechISBioTech.org

AMP Meeting Focuses on the Need for Standardization in the Global MarketThe Association for Molecular Pathology (AMP) held its 2016 meeting on November 10-12, 2016 at the Charlotte Convention Cen-ter in Charlotte, NC. The program consisted of an AMP reference materials (RM) forum, a science educator’s workshop, a molecular pathology outreach course, and a scientific program to name a few.

The AMP 2016 International Showcase fo-cused on the need for testing consistency and standardization of molecular testing world-wide.

Several CSP members were in attendance, with Liz Kerrigan from the ATCC presenting a workshop on standards, and the senior di-

rector of the ATCC Standard Resource center, Dr. Maryellen de Mars, speaking on advanc-ing standards for clinical diagnostics. For more information:https://www.amp.org/meetings/2016/index.cfm

Workshops & Presentations

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KAVI User WorkshopKAVI held its fifth annual Kavi User Group Workshop on Wednesday November 30th, 2016 at the American Society of Clinical On-cology Conference Center in Alexandria, VA. The user group workshop provided an oppor-tunity to share company vision, product prog-ress, and development plans that would assist the SDO users. The interactive setting also al-lowed 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 stan-dards development.

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

ANSI/ATCC ASN-0002 Revision and ReaffirmationANSI/ATCC ASN-0002 Authentication of Human Cell Lines: Standardization of STR Profiling is scheduled for review and revision in 2017.

Standards approved as American National Standards must be kept current and relevant by means of a timely review. Following the review, there are 3 choices--- the standard will either be revised, reaffirmed (mean-ing no changes are needed), or withdrawn. Standards that are no longer relevant, are obsolete, or expired will be withdrawn per the SDO Policy and Procedure manual, sec-tion 20.0 titled CRITERIA FOR APPROVAL AND MAINTENANCE OF AN AMERICAN NATIONAL STANDARD. This continuous maintenance is also addressed in the ANSI Essential Requirements section 4.7.1 where periodic maintenance of each American Na-tional standard must be revised or reaffirmed at 5 years, or else it may be recommended for withdrawal.

Members of the original workgroup were contacted in Q4 of 2016 to see who would be willing and available to assess the exist-ing standard and recommend a disposition. A project initiation notification form (PINs) was submitted to ANSI, and an announcement posted in the ANSI Standards Action to the public.

To give you a very brief background, ANSI/ATCC ASN-0002 Authentication of Human Cell Lines: Standardization of STR Profiling received ANSI approval on January 25, 2012. It focused on a standardized procedure for au-thentication and identification of human cell lines using STR profiling. This standard con-tains a method for DNA extraction, a method for STR profiling, data analyses, quality con-trol of the data, interpretation of the results, and how to use a searchable public database.

Social Media

The 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 informa-tion of interest.

Join our LinkedIn network!

Workshops & Presentations

(continued)

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World Standards Week 2016The ATCC SDO joined the global standardization community in cel-ebrating World Standards Week, which began on October 24th and ran through October 28, 2016. Meetings and related activities were held at ANSI Headquarters and the FHI 360 Conference Center in Washington DC. Amidst the workshops and meetings, was the U.S. Celebration of World Standards Day on October 27, 2016. This year’s festivities recognized the important role of stakeholders across the standards com-munity from business leaders, industry, academia, and government.

World Standards Week is held annually, and pays tribute to the thou-sands 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 standardiza-tion community were in attendance throughout the week-long series of events. For more information and to view the full list of activities, go to https://ansi.org/meetings_events/wsw16/wsw.aspx?menuid=8

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Frequently Asked Questions

1. Question: What is Cell Line Authentication?

An authentic cell line is a cell line of a specific species that retains its unique functionality and characteristics. Cell line authentication can be said to be a set of tests intended to demonstrate that a cell line is what it is purported to be – comprised of the same cells as the original primary or continuous cell culture from which the cell line was derived, free of cross-contaminating inter-species cells, and if possible, intra-species cell types, of expected morphology, ploidy, and phenotype under a given set of culture conditions. The authenticity of the cell line may be compro-mised due to species misidentification, sub-optimal growth conditions and contamination with adventitious agents

2. Question: What are some of the technologies being used to authenticate cell lines?

Traditional methods like HLA typing and karyotyping, molecular methods like

CO1 Barcoding, STR Profiling, SNP Profiling, and Next Generation se-quencing, light and transmission electron microscopy, and flow cytometry.

NOTE: For Intra-species identification in human cell lines, one can use STR analysis and or SNP analysis. For interspecies identification of ani-mal cell lines, one can use CO1 barcoding, and or Karyotyping.

3. Question: When should cell lines be checked/authenticated?

▪ On arrival at the facility, if not obtained from a reputable source/ repository with a certificate of authenticity

▪ When establishing a cell bank or cell stock prepared at your facility

▪ If marked phenotypic or morphological changes are observed during routine culture

▪ All of the above should be done prior to publishing results based on the cell line

4. Question: Where can I learn more?

Short tandem repeat profiling: part of an overall strategy for reducing the frequency of cell misidentification.Nims RW, Sykes G, Cottrill K, Ikonomi P, Elmore E. In Vitro Cell Dev Biol Anim 46:811-819, 2010. PMID: 20927602

ANSI/ATCC ASN-0002-2011 (2012) Authentication of human cell lines: Standardization of STR profiling. ANSI eStandards Store.

Short tandem repeat profiling provides an international reference standard for human cell lines.Masters JR, Thomson JA, Daly-Burns B, Reid YA, Dirks WG, Packer P, Toji LH, Ohno T, Tanabe H, Arlett CF, Kelland LR, Harrison M, Vir-mani A, Ward TH, Ayres KL, Debenham PG.Proc Natl Acad Sci U S A. 2001 Jul 3;98(14):8012-7.

Recommendation of short tandem repeat profiling for authenticating human cell lines, stem cells, and tissues.Barallon R, Bauer SR, Butler J, Capes-Davis A, Dirks WG, Elmore E, Furtado M, Kline MC, Kohara A, Los GV, MacLeod RA, Masters JR, Nardone M, Nardone RM, Nims RW, Price PJ, Reid YA, Shewale J, Sykes G, Steuer AF, Storts DR, Thomson J, Taraporewala Z, Alston-Roberts C, Kerrigan L.In Vitro Cell Dev Biol Anim. 2010 Oct;46(9):727-32. doi: 10.1007/s11626-010-9333-z

Authentication of Human Cell Lines by STR DNA Profiling Analysis.Reid Y, Storts D, Riss T, Minor L.In: Sittampalam GS, Coussens NP, Nelson H, Arkin M, Auld D, Austin C, Bejcek B, Glicksman M, Inglese J, Iversen PW, Li Z, McGee J, Mc-Manus O, Minor L, Napper A, Peltier JM, Riss T, Trask OJ Jr., Weidner J, editors. Assay Guidance Manual [Internet]. Bethesda (MD): Eli Lilly & Company and the National Center for Advancing Translational Sci-ences; 2004-. 2013 May 1.PMID: 23805434

Reproducibility: changing the policies and culture of cell line au-thentication.Freedman LP, Gibson MC, Ethier SP, Soule HR, Neve RM, Reid YA.Nat Methods. 2015 Jun;12(6):493-7. doi: 10.1038/nmeth.3403. No ab-stract available. PMID:26020501

Characterization and authentication of cancer cell lines: an overview.Reid YA.Methods Mol Biol. 2011;731:35-43. doi: 10.1007/978-1-61779-080-5_4. Review. PMID:21516396

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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 [email protected]. 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.

© 2016 American Type Culture Collection. The ATCC trademark and trade name are owned by the American Type Culture Collection.

Dr. Donna McAllisterResearch AssociateDwinell LabDepartments of Microbiology and Biophysics Medical College of Wisconsin

Matthew Winans StudentWest Virginia University

Dr. Kai LiMarketing ManagerDonghua UniversityChina

Suman GiriStudentPokhara Nepal

Balsam ShawkyBiologistATCC

Dr. Weihong WangTechnology Development MangerEurofins Lancaster Laboratories

Dr. Joe MarcinoAquatic PathologistArizona Game and Fish

Mousumi DasMicrobiologistRabon ChemicalsIndia

Dr. Li LiTechnical ManagerGene CreateChina

ATCC SDO WelcomesNew CSP Members

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