February 22 - 24, 2017

Wednesday - Friday

Germantown, MD

The Bioscience Education Center

9:00am-5:00pm

1 Hour Lunch Break

This hands-on three-day workshop is designed for scientists involved in laboratory research using cell cultures and or antibodies.

ADDRESSING

Cell Line Authentication • Cross Contamination • Mycoplasma Detection • Antibody Characterization/Validation

Identified as significant contributors to the current quality-control challenges in cell culture-based research*, this workshop will address methods and practices to ensure reproducibility and good cell culture practice (GCCP). Benefits also will extend to satisfy the new NIH grant requirements as well as those of publication outlets.

*NIH Workshop on Reproducibility in Cell Culture Studies, September 28-29, 2015

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Lecture and Hands-on Interactive Training
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Team taught by active researchers
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Comprehensive binder containing workshop material
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Space limited to 24 participants
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Registration Fee: $895

Course Director

Roland M. Nardone, Ph.D.
Professor Emeritus
Catholic University of America

Cross Contamination/Misidentification: Consequences Detection and Prevention; GCCP

Cell Line Authentication: Authentication of Human Cell Lines by STR Profiling Including Data Analysis; NIST STR Standards; Authentication of Non-Human Cell Lines: Isozyme Analysis; CO1 Multiplex Assays, STR Profiling

Mycoplasma Contamination: Severity, Causes, Effects and Prevention; Testing Regimens: Efficacy of Various Mycoplasma Tests; FDA Gold Standard, WHO Protocol, Culture test, DNA stain (Hoechst 33258), and PCR Tests; Rescue of Valuable Contaminated Cell Lines; Alterations in Gene Expression; Detection of Transcriptisome Contamination Studies (RNA-seq); How to Detect Contaminated Transcript files using NGS

Antibodies: Overview: Specific Issues Associated with the Use of Antibodies Including Antibody Origins (Mabs vs. Polyclonal Abs), Cross-Reactivity, Genetic Drift of Hybridomas; Care and Maintenance of Hybridomas; Consequences of Using “Bad” Antibodies; Purity of Antibody Preparations: Antibody Contamination, Standard Methods for Detection (SDS-PAGE, HPLC), in vivo vs. in vitro; Purification of Antibodies Bias: Changing Isotype Distribution/Profile of Polyclonal Antibodies Depending on Purification Method; Antibody Validation: Addressing the Difference Between Validation and Batch-to-Batch Variations; Isotype Confirmation: ELISA Based Checks for Contamination with Other Ab; Western Blot: Advantages and Limitations; Immunoprecipitation followed by SDS-PAGE: Overcoming WB-limitation of Many Mabs (how does IP work, how to “adapt” it to use of mabs); Flow Cytometry; Blocking studies: Using Competing Antibodies, Blocking Peptides/Proteins to Verify Specificity

Course Director

Dr. Roland M. Nardone is Professor Emeritus at the Catholic University of America.

Throughout most of his 60 year research career he used cell cultures as experimental tools for a variety of studies including cell cycle kinetics, nucleolar function and structure, in vitro toxicology, and microwave effects on gene expression. During that time he also served as Director of the Center for Advanced Training in Cell and Molecular Biology. During the past decade he devoted his attention to resolution of the problem caused by the rampant use of misidentified and cross-contaminated cell lines, an initiative that culminated in policy changes regarding grant applications and manuscripts submitted for publication. He is currently addressing remediation of a similar problem, namely the use of untested and mycoplasma contaminated cell lines for research. Dr. Nardone is a member of the International Cell Line Authentication Committee (ICLAC) and the ASN-0002 Committee for Authentication of Human Cell Lines: Standardization of STR Profiles.

WLeitnerWolfgang Leitner, Ph.D.

Dr. Wolfgang Leitner the Chief of the Innate Immunity Section at the National Institute of Allergy and Infectious Diseases (NIH). He has been teaching flow cytometry for 25 years. He previously worked at the National Cancer Institute where he developed various flow cytometric protocols for the evaluation of experimental cancer vaccines.

The Bioscience Education Center

Montgomery College
20200 Observation Drive
Germantown, MD 20876