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Epithelial Organoids and 2D Monolayer “EpCulture” Services Offered by the Advanced Analytics Core
Organoid technologies have transformed in vitro GI tract research, but they pose unique challenges due to their non-traditional culture methods. Organoids are grown in 3D matrigel patties that require more difficult culture techniques than standard 2D methods, and their spherical structures are challenging to evaluate by microscopy. To address these challenges, investigators at UNC have pioneered 2D monolayer culture methods that use the same initial starting material as 3D organoid cultures (typically epithelium in the form of small intestinal or colonic crypts). 2D monolayers are more convenient to work with since they can be passaged and cryopreserved using standard methods. The UNC Advanced Analytics Core offers 2D monolayer and 3D organoid culture services, AKA “EpCultures”, to interested UNC/NCSU investigators. We also offer specialized conditioned media to facilitate both 2D and 3D organoid culture.


EpCulture Services (Pricing Determined after Consultation)
:
  1. Conditioned Media
    1. L-WRN (L-cell secreted Wnt3a, R spondin 3, and Noggin) Conditioned Media
    2. Expansion Media (maintains intestinal stem cells; available upon request)
    3. Differentiation Media (no Wnt3a, R spondin 3, and Noggin; available upon request)
    4. More info available here
  2. Human Intestinal Stem Cells (hISCs)
    1. Provision of dissociated hISCs with option of which region the investigator would prefer: Duodenum, Jejunum, Ileum, Ascending Colon, Transverse Colon, Descending Colon
  3. Multi-Well Cell Expansion
    1. Custom cell culture expansion and cryo-preservation of EpCultures using established mouse models or human samples, either provided by users or procured from AAC
    2. Expansion onto Transwell or Collagen plates
  4. 2D/3D organoid generation (dissociating crypts from tissue)
    1. Isolation of intestinal and colonic stem cells (ISCs) from patient biopsies, tissue resections, and mouse models
  5. EpCulture Gene-Editing
    1. Gene-editing by CRISPR or PiggyBack Transposition (consultation required)
  6. Experimental Design Consultation
    1. With expertise in hISCs provided by AAC
  7. Training
    1. ISC maintenance
    2. Passaging and freezing
    3. Extracellular Matrix (ECM) establishment
    4. Gene editing (consultation required)

Submitting Orders:

Please email Quinn Fong (xequinn@med.unc.edu) regarding any service requests or general inquiries


AA Core MBRB Laboratory

Medical Biomolecular Research Building (MBRB)

Room 4331

111 Mason Farm Rd

Chapel Hill, NC 27599