Lesson 1Selecting in vitro models: organoids, co-culture systems, 2D vs 3D scaffolds and disease-relevant readoutsThis section discusses selection and design of in vitro models, including 2D cultures, 3D scaffolds, organoids, and co-culture systems, with emphasis on microenvironmental cues, disease-relevant readouts, throughput, and alignment with in vivo findings.
2D monolayer versus 3D culture systemsOrganoid models for organ-specific diseasesCo-culture and immune–stromal interactionsBiomaterial scaffolds and matrix mechanicsAssay readouts linked to in vivo outcomesThroughput, scalability, and assay robustnessLesson 2Small animal models: mouse and rat disease models relevant to chosen indication and translatability limitsThis section focuses on small animal models, mainly mouse and rat, describing disease models relevant to regenerative indications, genetic tools, immune backgrounds, practical advantages, and key limitations for predicting human safety and efficacy.
Mouse versus rat: strengths and weaknessesImmunodeficient and humanized rodent modelsCommon disease models for key indicationsModel validation and relevance to humansLimitations in scale, immunity, and lifespanBridging data to large animals and clinicsLesson 3Dosing strategies and kinetics: cell dose-ranging studies, biodistribution, and persistence analysisThis section explains dosing strategies for cell therapies, including dose-ranging designs, single versus repeat dosing, route-dependent dose adjustments, biodistribution mapping, persistence and engraftment analysis, and modeling exposure–response relationships.
Design of dose-escalation and range-findingSingle versus multiple dosing regimensLabeling cells for tracking in vivoBiodistribution assessment and organ tropismPersistence, engraftment, and clearance kineticsExposure–response and safety margin modelingLesson 4Large animal models: pig, sheep, non-human primate choices for scale-up and functional similarityThis section examines large animal models such as pig, sheep, and non-human primates, focusing on anatomical and physiological similarity, immune context, housing and welfare, scaling of devices and doses, and when large models are essential for translation.
Criteria for choosing large animal speciesPig models for cardiovascular and liver repairSheep models for orthopedic and spine studiesNon-human primate models and ethicsScaling of devices, doses, and proceduresHusbandry, welfare, and regulatory oversightLesson 5Outcome measures: histology, functional assays, imaging modalities (MRI, PET, bioluminescence), molecular biomarkers, and behavioral testsThis section details how to select and validate outcome measures, including histology, functional assays, imaging modalities, molecular biomarkers, and behavioral tests, emphasizing sensitivity, specificity, quantification, and relevance to human clinical benefit.
Histological scoring and morphometric analysisIn vitro and ex vivo functional assaysMRI, PET, and optical imaging strategiesMolecular biomarkers and omics readoutsBehavioral tests and functional performanceValidation, standardization, and assay QCLesson 6Delivery routes and device-assisted delivery: intravenous, intra-organ, intra-articular, biomaterial scaffolds, and catheter-based techniquesThis section reviews delivery routes and device-assisted methods for cell and gene-modified products, including intravenous, intra-organ, intra-articular, scaffold-based, and catheter techniques, highlighting targeting efficiency, safety, and technical feasibility.
Intravenous and intra-arterial deliveryDirect intra-organ and parenchymal injectionIntra-articular and intradiscal approachesBiomaterial scaffolds and hydrogelsCatheter-based and image-guided deliveryProcedure-related risks and mitigationLesson 7Study design principles: hypotheses, controls, randomization, blinding, sample size calculation, and statistical endpointsThis section introduces rigorous preclinical study design, from defining hypotheses and selecting controls to randomization, blinding, sample size calculation, and statistical endpoints, aiming to reduce bias and improve reproducibility and regulatory acceptance.
Formulating testable mechanistic hypothesesChoice of control and comparator groupsRandomization schemes and allocation concealmentBlinding of investigators and outcome assessorsSample size and power calculationsPrimary, secondary, and exploratory endpointsLesson 8Cell sourcing, manufacturing and GMP considerations for preclinical batches: isolation, expansion, cryopreservation, and release criteriaThis section covers sourcing and manufacturing of preclinical cell batches, including tissue selection, isolation, expansion, cryopreservation, quality control, GMP compliance, and release criteria to ensure consistent, safe, and well-characterized products.
Donor selection and tissue procurementCell isolation and early characterizationExpansion, passaging, and senescence controlCryopreservation methods and thaw recoveryGMP documentation and batch recordsRelease testing and specification setting
