Gastrointestinal Organoid Derivation and Culture

Core Scientists: David Breault, MD, PhD, Camilla Richmond, MD                  
Core Technicians: Eric Stas, Elizabeth Howe Vera                                        Fusing Organoid       
 
Services                                                   

This service provides in-depth protocols, hands-on training, biologics, and ongoing support for the derivation and culture of gastrointestinal organoids/enteroids/mini-guts from stomach, small intestine, or colon of mouse or human.

The L-WRN cell line (L cells expressing Wnt3a, R-spondin3, and Noggin) is maintained and aliquots of conditioned media are available.

The R-Spondin cell line (293T cells expressing mouse R-Spondin1) is maintained and aliquotes of conditioned media are available.

Fee Schedule

Organoid Services HDDC Member Non-Member
Hands-on training no charge $100/h
Consultation no charge $100/h
Corning® Matrigel® Growth Factor Reduced (GFR) Basement Membrane Matrix - Phenol Red-Free, LDEV-Free, 10mL $300 $320
(Limit 15 vials per year, additional vials will be charged as non-member rate)    
Recombinant Murine Noggin (100 micrograms) $375  
L-WRN Conditioned media (500 ml) $400 $550
L-WRN Conditioned media (Antibiotic Free) (500 ml)   $600
(Limit 5 bottles of L-WRN per year, additional bottles will be charged as a non-member rate)    
R-Spondin Conditioned Media (250 ml) $145 $260
Invitrogen EVOS FL Auto 2 Imaging Service no charge  
              Smaller volumes of these conditioned media are available at a prorated rate.      

Logistics:

Investigators are trained in organoid culture by Core scientists (David Breault, MD, PhD and Camilla Richmond, MD) and staff. The Core helps investigators to establish this technology within their own labs, by providing the resources for a “pilot experiment” using primary tissue provided by the investigator or the core as needed. Methods include isolation of crypts/glands from primary tissues, suspending them in Matrigel, culture in defined or conditioned media, and monitoring organoid health. Methods to perform viral transduction, organoid freeze down, fluorescent imaging of live cultures and post-fixation histological applications are also available. Consultation is provided to HDDC members (at no charge).

Quality Control:

L-WRN conditioned media used for organoid generation are routinely assayed for Wnt-3a activity.