August 22 - 26, 2016

Monday - Friday

Germantown, MD

The Bioscience Education Center

9:00am-5:00pm

1 Hour Lunch Break

This rigorous five day program is ideal for basic research and translational biology scientists who are looking for a balanced theoretical vs. hands-on introduction to reprogramming utilizing non-integrating systems, culturing and differentiating human iPSCs and different CRISPR-based tools for gene editing in iPSCs.

SOLD OUT

  • Lecture and Hands-on Interactive Training

  • Team taught by active researchers

  • Comprehensive binder containing workshop material

  • Space limited to 15 participants

  • Registration Fee: $1,095

Course Co-Director

Joseph Bressler, Ph.D.
Research Scientist, Kennedy Krieger Institute

Course Co-Director

James Kehler VMD, PhD
Director of Scientific Alliances, MTI-GlobalStem

Presentations in the reprogramming, gene-editing and iPSC differentiation fields

Strategies for optimizing delivery of CRISPR/Cas9 DNA, RNA and Ribonucleoprotein complexes into iPSCs

Techniques  for isolation of clonal iPSC lines

Wet labs designed for all aspects of daily propagation and use of iPSCs:

  • Isolating colonies

  • Passaging and expanding clonal lines

  • Banking to characterization and basic differentiation

  • Meso/endodermal and neurectodermal lineages

Lectures:

  1. Fundamentals of Pluripotency and iPSC Culture
  2. Fundamentals of Reprogramming
  3. Comparison of Non-Integrating Reprogramming Systems

Human iPSC Culture Activities:

  • Preparing Culture Media and Substrates
  • Passaging of iPSCs
  • Thawing iPSCs

Reprogramming Activities:

  • Plating Human Fibroblasts for Reprogramming
  • Detection of Live Pluripotency Markers
  • Manual Isolation of Primary iPSC Colonies
  • Single Cell Cloning of iPSCs

Lectures:

  1. Reprogramming with Episomal Plasmids
  2. Reprogramming with Sendai Viral Vectors
  3. Reprogramming with RNA and Trouble Shooting

Human iPSC Culture Activities:

  • Daily Care and Review of Human iPSC Cultures
  • Passaging iPSCs
  • Plating iPSCs for Gene Editing
  • Generating Embryoid Bodies

Reprogramming Activities:

  • Transfection of Episomal Plasmids
  • Transfection of mRNAs
  • Transduction of Sendai Viral Vectors into Patient Fibroblasts

Lectures:

  1. Fundamentals of Gene Editing
  2. Gene Editing with CRISPR/Cas9
  3. Optimizing Gene-Editing Experiments in iPSCs

Human iPSC Culture Activities:

  • Daily Care and Review of Human iPSC Cultures
  • Passaging iPSCs
  • Cryo-Preserving iPSCs
  • Plating iPSCs for gene-editing
  • Plating iPSCs for directed differentiation in monolayer cultures
  • Plate NSCs

Reprogramming Activities:

  • Assessing Transfection/Transduction Efficiency by Epifluorescence

Lectures:

  1. Meso/Endodermal Differentiation
  2. Neurectodermal Differentiation
  3. Optimizing iPSC Workflows

Human iPSC Culture Activities:

  • Daily Care and Review of Human iPSC Cultures
  • Thawing cryopreserved iPSCs from Day 2 to check viability
  • Initiate Monolayer Differentiation protocols

Reprogramming Activities:

Transfect CRISPR/Cas9 editing complexes into iPSCs and NSCs:

  • DNA Delivery of Cas9-2A-GFP sgRNA plasmids
  • Co-delivery of Cas9 mRNA+crRNA/tracrRNA
  • Ribonucleoprotein co-delivery of Cas9 protein with gRNA or crRNA/tracrRNA

Lectures:

  1. Optimizing HDR and Homologous Recombination
  2. Genomic Analyses of Gene-Edited iPSCs
  3. Final Question and Answer Session

Human iPSC Culture Activities:

  • Review of Human iPSC Cultures
  • Thawed iPSC cultures
  • Embryoid Bodies
  • Monolayer Differentiation cultures

Reprogramming Activities:

  • Review transfection/transduction reprogramming cultures
  • Review picked iPSC colonies and single cell clones

Gene Editing Activities:

  • Assessing Transfection Efficiencies

Course Co-Director

Dr. Joseph Bressler is a research scientist at Kennedy Krieger Institute. He is also an associate professor of environmental health sciences at the Bloomberg School of Public Health at Johns Hopkins University.

Dr. Bressler received his bachelor's of science degree in biology from the State University of New York at Stony Brook in 1973 and his doctoral degree in physiology from Rutgers University in 1978. His post-doctoral training at UCLA was in neurosciences, where he studied the involvement of glial cells in response to toxic agents. After his post-doctoral training, Dr. Bressler continued his studies on glial cells at the National Institutes of Health, Bethesda, MD. Dr. Bressler has been a research scientist at the Kennedy Krieger Institute since 1988.

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Course Co-Director

James Kehler VMD, PhD is a comparative stem cell biologist who thrives on developing productive collaborations to translate scientific discoveries into transformative products. He trained at the University of Pennsylvania, where he received his VMD in 2002, and PhD in Cell and Molecular Biology in 2004. James has worked as a visiting researcher at the National Institutes of Health for over 10 years, where he and his collaborators at NEI, NCI, NINDS, NIDDK and NIAAA developed animal and stem cell-based models of human diseases. He has run workshops on reprogramming both at the NIH, as well as internationally. James has worked and consulted for several stem cell companies from product development and management to directing custom reprogramming and gene-editing services. James recently joined MTI-GlobalStem, as Director of Scientific Alliances to foster collaborative research projects with academic, biotech and pharmaceutical partners.

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Richard Josephson, Ph.D. is a Senior Scientist with MTI-GlobalStem. He is primarily responsible for developing products and methods for neurobiology, including human iPS-derived neurons. He has been tasked with training and supervision for all primary neural cell production. He has been lead author of several papers and reviews on authentication and standards for human embryonic stem cells and stem cell therapies. Previously, Dr. Josephson was a research scientist in the Stem Cell Center at the American Type Culture Collection. Dr. Josephson conducted post-doctoral research on stem cells of the central nervous system in the National Institute of Neurological Disorders and Stroke at NIH. He received his Ph.D. in cell and molecular biology from the Massachusetts Institute of Technology.

The Bioscience Education Center

Montgomery College
20200 Observation Drive
Germantown, MD 20876