CORE C: INTEGRATED PHYSIOLOGY


Director, Nicholas C. Zachos, Ph.D.
nzachos1@jhmi.edu
410-614-0128


Associate Director, Ming Tse, Ph.D.
mtse@jhmi.edu
410-955-9681

GOALS

The Integrated Physiology Core provides multiple services and serves as a centralized source for equipment and reagents to enhance studies of epithelial biology in the gastrointestinal tract.

SERVICES

  • Murine and human gastrointestinal organoids
  • The Integrated Physiology Core provides training for establishing, expanding and maintaining mouse and human enteroid cultures. Core C instructs Center investigators how to isolate intestinal crypts from mouse and human tissues (i.e. biopsies or resections), seed into the 3D matrix (i.e. Matrigel), propagate and split cultures, cryopreserve cultures (to establish their own biobanks), and recover cultures from 3D for biochemical studies. The service allows Center investigators to develop this model within their own laboratories. The Core also prepares conditioned media containing the critical growth factors for investigators who study these organoid cultures. Fresh conditioned media is prepared weekly and provided to the Research Base on a fee-for-service basis at >90% cost savings.
  • Histology
  • The Core can process (fresh or frozen), embed (paraffin or OCT), section and stain tissues samples, including animal models and humans. The Core offers a variety of stains including hematoxylin and eosin, Masson’s Tri-Chrome Blue, Periodic Acid Schiff, Von Kossa, and Sirius Red. Paraffin and frozen sections may also be archived by the Core for future studies or collaborations.
  • Ussing chamber/Voltage clamp
  • The Core provides equipment and training for the measurement of active ion transport in cultured polarized epithelial cell models, ex vivo mouse and human intestine and human enteroid monolayers. Dr. Tse provides training for the equipment and advice for experimental design, including methods of calculation, quality control, and data interpretation. Applications include measurement of short circuit current, trans-epithelial resistance (TER), dilution potential, and paracellular dextran flux.

EQUIPMENT

• Physiological Instruments 6-Channel Voltage Clamp Connected to Easy Mount 6-Channel Ussing Chamber System
• World Precision Instruments EVOM2, Epithelial Voltohmmeter for TER in 24-well and 96-well formats

Multiplex Cytokine ELISA
The Core has established a multiplex ELISA system capable of analyzing various cytokines and/or chemokines simultaneously from the same serum or tissue samples obtained from mice and/or humans.

Bio-Rad 200 Plex
• Luminex-based multiplex ELISA systems

TRAINING

The Core provides extensive training for establishing and maintaining murine and human gastrointestinal stem cell-derived organoids.

FEES

  • Organoid training: $500 / session.
  • Weekly prepared enteroid media: $3 / ml (Full Members); $2 / ml (Associate Members)
  • Histology: $4.50 / specimen processing/embedding; $2 / unstained slide; $2.50 / stained slide.
  • Luminex Multi-plex ELISA training: $100 / session.
  • Luminex Multi-plex ELISA use: $40 / plate.

NEW SERVICE – 2019

Emulate “Gut-on-a-chip” microfluidic device automated incubation chamber