Functionally Characterized Human PBMCs

An Improved In Vitro Model of Human Immune Response


Abstract: Human Peripheral Blood Mononuclear Cells (PBMCs) contain many of the functional cell types of immune system, and are an ideal model to study the human immune response in vitro. Currently, PBMCs are being used in basic and clinical research areas including basic immunology and cell biology, infectious diseases, vaccine development, tumor immunology, and drug discovery. PBMC are also widely used for monitoring disease progression, designing personalized approaches to treatment, and predicting the occurrence of treatment-associated adverse events. The majority of PBMC applications rely on highly complex cell-based assays; however, phenotypic and functional variability of human PBMCs makes the development and validation of these assays difficult. Additionally, variations in cell collection and cryopreservation protocols can further affect PBMC properties. In this presentation, we demonstrate that pre-screening characterization of lot-specific functional activity of PBMCs is extremely useful for selecting cells to address individual experimental goals.

Key Points:

  • PBMCs play a critical role in modern biomedical research; however, the functional activity of these cells is highly variable
  • Many factors affecting PBMC variability, including genetic diversity and environmental pressure, are outside of researchers’ control
  • Conducting pre-screening and establishing PBMC functional activity profiles offers an attractive solution to address PBMC variability

Carbapenem-resistant Enterobacteriaceae

A Growing Superbug Population


Abstract: The discovery of antibiotics in the early twentieth century has revolutionized the treatment of infectious diseases, saving millions of lives and easing the suffering of many. However, as the structure and function of antibiotics have evolved through the efforts of biotech and pharma companies, prokaryotic species are evolving in parallel, fashioning novel and effective methods to avoid therapeutic killing. In the last several decades, this concern has become more pronounced with the emergence of multidrug-resistant organisms in both community- and hospital-acquired infections, resulting in increased morbidity, mortality, and health-care expense. In this presentation, we will discuss the emergence of multidrug-resistant infections with a particular emphasis on the emergence and global spread of carbapenem-resistant Enterobacteraeae strains.

Key Points:

  • Multidrug-resistant strains are an emerging problem throughout the world
  • ATCC acquires, authenticates, and distributes clinically relevant multidrug-resistant strains that are essential to the scientific community
  • KPC, NDM, and OXA strains are now available at ATCC

The ATCC Story

A Ninety Year Celebration


Abstract: ATCC celebrated its 90th anniversary in 2015 and in recognition of that milestone this presentation tells the remarkable story of ATCC’s evolution as an organization and its contribution to developments in life sciences for nearly a century. Established in 1925 as the American Type Culture Collection by scientists for scientists, it has become an international resource for life science research and development. Dependent on financial subsidies for more than 75 years, ATCC has since achieved independence as a fully self-sustaining non-profit organization. ATCC is now investing in research and development, and in 2012 was able to establish and fully fund the Global Biological Standards Institute (GBSI).This is the story of that amazing transformation and the role of ATCC in the evolution of the life sciences.

Key Points:

  • ATCC was founded by scientists for scientists; it is a science-based organization
  • ATCC is now fully self-sustainable after dependence on government subsidies and private financial support for more than 75 years
  • ATCC contributes to global science and health initiatives by providing biological resources and related services

Abstract: Cryopreservation is the use of very low temperatures to structurally preserve intact living cells and tissues. Normally, the freezing of water in cells causes catastrophic damage to cellular structure by physical damage of ice formation and increased imbalance of solutes. Cryopreserving cells with the proper cryoprotectants and techniques will maximize viability of cells for cell culture. This webinar presentation will discuss best practices for cryopreservation focusing on determining optimal freezing rates and cryoprotectants, selecting proper containment units, managing a biorepository, and handling cells post-thaw. Emphasis will be placed on reviewing time-proven techniques while introducing newer innovative approaches to maximize reliability in cryopreservation for modern day cell culture.

Key Points:

  • For the most accurate administration of DMSO, use Serum Free Cell Freezing Media already formulated with exactly 10% DMSO
  • A controlled cooling rate (1 – 3°C /min) is necessary to maximize cell viability when freezing down cell lines
  • The CoolCell® is a reliable and consistent cryo-container

A Tale of 3 Mummies

A Microbiome Analysis of Life in the Peruvian Andes 1,000 Years Ago


Abstract: The natural mummification process is a rare and unique process resulting from low temperatures and oxygen levels, and dry weather conditions. In the present study, we characterized the gut microbiome of three pre-Columbian Andean mummies using 16S rRNA gene high-throughput sequencing and metagenomics to understand the preservation and evolution of commensal and pathogenic microorganisms, antimicrobial resistance genes, diet, and the metabolic processes during the natural mummification of the human gut.

Key Points:

  • 16S rRNA gene high-throughput sequencing and metagenomics can be used to understand changes to the human microbiome
  • Evaluation of microbial populations from mummified remains can provide valuable insight into the medical and cultural aspects of ancient people
  • Ancient DNA must be suitable for investigation and should be evaluated through damage analysis studies
  • Quality control of the metagenome analysis process is key to ALL studies. It is particularly important in ancient DNA studies to ensure that the results are reflective of the actual sample, not an artifact of DNA extraction, PCR, sequencing, and/or analysis

Solving Identity Crisis in Animal Cells

Best Practices with DNA Barcodes and STR Analysis


Abstract: Animal cell lines are important in vitro systems and tools for scientists in diverse disciplines such as basic cell biology, genetic mapping, gene expression, and gene therapy. Animal cell line identity, which is a crucial first step in cell line authentication, is frequently underappreciated and ignored by most research scientists. Over the years numerous cell lines have been shown to be misidentified due, in part, to poor techniques and inadequate authentication protocols. It is estimated that the financial loss incurred by misidentified cell lines is in the millions of dollars. Animal cell line identity now requires a comprehensive strategy that employs several complementary technologies. Technological advances have given rise to improved capabilities. An overview of these technologies to include STR analysis for human cell line identity and CO1 for animal cell line identity will be presented.

Key Points:

  • Cell line authentication is critical to meet funding, quality control, and publication requirements
  • Short tandem repeat (STR) profiling is an accepted form of authentication that differentiates human cell lines
  • DNA barcoding complements STR profiling by providing species-specific identification

Neural Progenitor Cells

Potent Models of Normal and Disease Neurobiology


In vitro neurological research presents many challenges due to the difficulty in establishing high-yield neuronal cultures as well as batch-to-batch consistency. Human induced pluripotent stem cells (iPSCs) have a high expansion capacity and can differentiate into neurological cells types; thus, these cells hold great promise for both regenerative medicine and drug discovery. This webinar will discuss neural differentiation and touch on its roles in investigating neurodegenerative diseases and screening therapeutics. Methods for generating large quantities of neural progenitor cells (NPCs) from human fibroblast-, CD34+-, and Parkinson’s disease-derived iPSCs will then be highlighted. Finally, the generation and function of NPC reporter cell lines, created using zinc finger nuclease gene editing technology, will be examined.

We often speak of the global technical community in the abstract, but accreditation to ISO 17025 makes each accredited laboratory feel part of the practical everyday operation of that community. With our ever-growing science and technologies, manufactured and agricultural materials, telecommunications, internet, cyber-security, weapons, explosives, drugs, medicines, and our homeland security operations and interdiction testing, we need to have confidence that materials are sampled well, tested appropriately, and that their measurement tools are well calibrated. The oversight and coordination of all calibration and testing is standardized and managed as best as possible through ISO, through ILAC, and in the everyday, through labs accredited to ISO 17025.

Phenotype MicroArray (PM) technology allows a biologist to test thousands of phenotypes of a cell line in a single experiment to gain a comprehensive overview of the metabolism, physiology, and pathway fluxes. It provides phenomic and metabolomic information that is complementary to genomic or proteomic analysis and often more easy to interpret and more useful. The PM technology platform is applicable to a wide range of cells including bacterial, fungal, or animal and enables metabolic analysis in the context of genotype-phenotype studies. For example, it can be used for (1) analyzing cells with mutations to determine the metabolic and physiologic effects of genetic differences, (2) studying and defining cell metabolism and metabolic regulation, (3) understanding the interplay of environment and hormonal signals on cell metabolism and physiology, (4) optimizing cell culture conditions in bioprocess development and optimization, and (5) examining and maintaining the stability of cell lines. Specific examples and discoveries will be presented to illustrate the many uses of this cell phenotyping technology.

The rapid identification of microorganisms is a great advantage for industrial and clinical laboratories. In the 150 year history of microbial identification, many techniques have been used to authenticate microorganisms, including biochemical analyses, sequencing, and functional testing. The techniques vary in turnaround time, complexity, accuracy, and specificity. The VITEK®-MS is a recent addition to the arsenal of identification technologies. It brings to the field a very simple user interface, rapid turnaround times, and flexibility in expanding a database to suit the user’s needs. In this webinar, we will discuss current methods of microbial identification and how VITEK®-MS compares to these methods with regard to complexity, turnaround time, sample preparation, and ease of interpretation.