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HUMANOID Services

The Phase Zero Approach

  • There is a "translational valley of death": The success rate for a new target identified to reach the next step in the drug discovery process is undeniable low and wrought with failure. This is because many of the pre-clinical models to date do not accurately recapitulate the human system, and assessment of efficacy typically waits until Phase III clinical trials.
  • HUMANOID is dedicated to build and validate pre-clinical human healthy and disease models as platforms for research and drug development, and enable early ex vivo testing for toxicity and efficacy.
  • With few exceptions, all tissues from human, mouse, embryonic, adult, diseased, normal, iPSCs, can be developed into organoids, and subsequently into disease models with a few network-guided tweaks.
  • Our ex vivo models recreate the complexity in vivo by making multi-dimensional organoids [co-culture techniques that capture the 3D of organoid tissues + microbiome + immune or non-immune cells]
  • In powerful synergy with iNetMed's precise AI-assisted target discovery, HUMANOID can build personalized models with patient tissue; together, they enable "precisionalized" medicine.
  • HUMANOID collaborates with an extensive network of clinicians, industry collaborators and basic scientists, both on and off campus.
  • HUMANOID's services include enabling the research community to access and experiment with human primary cells, organoids, enteroid derived monolayers, derivatives of organoids, support sponsored research projects, provide consultation, offer formal and informal education on organoid research and more.
  • HUMANOID is located in the George Palade Laboratories (GPL) for Cellular and Molecular Medicine [Room 238].

 

Frequently Used Applications of Our Core

Patient Derived Organoids:

  1. Treatments with therapies or stressors. 

  2. RNA, DNA and supernatant isolation/collection 

  3. Proliferation:

    1. Ki67 (IF, Flow)
    2. BrdU (ELISA)
    3. MTT

  4. Apoptosis:

    1. AnnexinV (IF)

  5. Cytotoxicity:

    1. LDH Assay
    2. Zombie Viability Kit
    3. LIVE/DEAD Viability Kit
  6. DNA Damage:
    1. Gamma-H2AX (IF, Flow)
    2. Oxidative DNA/RNA damage (ELISA Plate based kit)
  7. Senescence:
    1. SA-β-gal
  8. Immunoflourescence:
    1. 8-chamber slide for confocal imaging and z-stack with 3D rendering if needed
    2. 96-well format (BioTek Cytation 10 Confocal Imaging Reader)
    3. FFPE slide (from histogel)

Monolayers Derived from PDOs: 

  1. Submerged (TC plate and transwell inserts) and Air-Liquid models
  2. RNA, DNA, and supernatant isolation/collection
  3. Barrier function (baselline and post therapy):
    1. TEER reading (24-well and 96-well  format)
    2. Paracellular Permeability (FITC-Dextran, Cascade Blue)
  4. Treatment with stressors or therapies 
  5. Co-culture systems with monolayers 
    1. Bacterial (probiotic and pathogenic)
    2. Viral 
    3. Human immune and mesenchymal cells
  6. Bioenergetics - Seahorse
  7. Senescence:
    1. SA-β-gal
  8. Immunoflourescence (transwell and TC plate)

Organoid Support

  1. Training modules (stem cell isolation, passaging, freezing, monolayers)
  2. Media
    1. Organoid:  Intestigro, Stomagro, Lungbrew
    2. Ancillary: Orga-Freeze, Tailor-2-Gro, Tissue-2-Go 
  3. Consultation

Trademarked Media Products

HUMANOID’s Computationally vetted organoid models:

  • Inflammatory Bowel Disease: Over 350 unique Crohn’s Disease and Ulcerative Colitis subjects; many lines come with corresponding myofibroblasts from the same subject. Selected lines have undergone comprehensive phenotype, genotype, and transcriptome analyses. All lines feature thorough metadata, encompassing patient demographics, clinicopathologic behavior, histology reports, treatment information, and, where applicable, patient response. 
  • Hereditary Cancer (Predisposition) Syndromes: Familial adenomatous polyposis (FAP), juvenile polyposis syndrome (JPS), Lynch syndrome (HPNCC), and Peutz-Jeghers syndrome (PJS). Includes repeat biopsies from the same subjects for longitudinal studies and multiple organ locations in some subjects, such as colon, small intestine, gastric, and esophageal tissues. All patient-derived organoids come with pathology reports and insights into their diagnosis-defining mutation. 
  • Foregut Diseases (pre-cancer): Barrett’s esophagus, eosinophilic esophagitis, gastric intestinal metaplasia, GERD, as well as healthy controls. 
  • Lung Inflammation, Fibrosis, and Cancer: Several unique subjects, some of whom have matching tumor and normal organoids. PDO lines have been extensively characterized for retaining all 5 epithelial cell types in culture, essential for modeling alveolar diseases. These lines are valuable for studying lung inflammation (COVID-19, bacterial), lung fibrosis, the effects of anti-fibrotic therapies using EVs and small molecules, and immunotherapy impact. Information includes smoker status but not tumor genotype. 
  • Cancer Patient-Derived Organoids (PDOs): Several cancer PDO lines, representing various cancers (colon, biliary, pancreas, gastric, and esophagus). Some of these are from the ATCC, the Broad Institute and Columbia University. These lines are available for researchers to conduct organoid-based cancer studies, emphasizing our motto, "We bank, so that you can avoid the hassle and just focus on your planned studies!". 

Recharge Core Services

  1. Equipment
  2. Media and Cocktails and starter kits [mouse or human or BOTH]
  3. Primary cells, DNA/RNA/lysate [mouse or human or BOTH]
  4. Consultation for experimental design and troubleshooting
  5. Organoid-based experiments [mouse or human or BOTH]
  6. Access to plate-based disease models for customized assays
  7. Training for students and fellows available to those in academic or industry

Contact Us

Courtney Tindle, M.S.
ctindle@health.ucsd.edu

 

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