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3D Cell Culture

Introductory 3D Cell Culture: From 2D Monolayers to Spheroids

XTech Academy in collaboration with CAVIGEN

Hands-on

Course Details

Overview

Step into the next generation of cell biology with our intensive 3-day hands-on workshop on 3D Cell Culture, designed for students, researchers, and professionals seeking practical experience with modern in vitro models used in biotechnology, cancer research, drug development, and regenerative medicine.

Traditional 2D cell culture systems often fail to accurately mimic the complexity of living tissues. In contrast, 3D cell culture models, including spheroids and microtissue systems, provide more physiologically relevant environments that better reproduce cell–cell interactions, tissue architecture, nutrient gradients, and drug responses observed in vivo. These advanced systems are increasingly becoming essential tools in translational research, toxicology, immunotherapy development, and precision medicine.

During this workshop, participants will gain both theoretical understanding and practical laboratory experience in the generation, handling, staining, imaging, and interpretation of 3D cell culture models. The course combines cell culture techniques, fluorescence microscopy, confocal imaging, live/dead staining, and experimental analysis in a highly interactive learning environment.

Participants will work directly with different 3D culture platforms, including hanging-drop systems, microcavity arrays, and specialized 96-well plates. Throughout the course, attendees will document and analyze their own experiments while learning how to troubleshoot common technical challenges associated with 3D cultures.

Learning Objectives

By the end of this workshop, participants will be able to:

  • Understand the biological advantages and limitations of 3D cell culture systems
  • Generate and maintain spheroids using different culture platforms
  • Perform live/dead staining and fluorescence-based analyses
  • Operate fluorescence and confocal microscopy systems
  • Evaluate and troubleshoot 3D culture experiments
  • Interpret imaging results and communicate scientific findings effectively
  • Understand current applications of spheroids, organoids, and organ-on-chip technologies in biomedical research

Target Audience

This course is ideal for:

  • Master’s and PhD students
  • Research assistants and laboratory technicians
  • Biotechnology and biomedical researchers
  • Scientists transitioning into advanced cell culture technologies
  • Industry professionals interested in drug discovery, cancer research, and tissue engineering

Course Program

Day 1: Introduction to 3D Cell Culture

Time Activity Format
8:00 – 8:15 Introduction & Course Overview Lecture
8:15 – 9:15 Introduction to BSL-2 Guidelines Lecture
9:15 – 10:00 Basics of 2D vs. 3D Cell Culture, Safety and Lab Organization Lecture
10:00 – 10:30 Cell Culture Refresh: Harvesting, Counting and Adjusting Cell Density Laboratory
10:30 – 12:30 Seeding 3D Cultures: Each participant seeds 1 Hanging-drop Plate (10 cm Petri Dish), 1 MicroSphere Array, and 1 Row of a faCellitate 96-Well Plate Laboratory
12:30 – 13:30 Lunch Break
13:30 – 14:00 History and Landscape of 3D Cell Culture: From Simple Aggregates to Organoids and Organ-on-Chip Systems Lecture
14:00 – 15:00 Block 1 (Split Groups) • Group A: Trypsinisation and Cell Counting Practice on Individual 2D Culture Flasks • Group B: Introduction to the Fluorescence/Confocal Microscope (Controls, Focus, Filters, Best Practices) Laboratory
15:00 – 15:15 Coffee Break
15:15 – 16:15 Block 2 (Groups Switch) • Group B: Trypsinisation and Cell Counting Practice on Individual 2D Culture Flasks • Group A: Introduction to the Fluorescence/Confocal Microscope Laboratory
16:15 – 16:45 Experiment Documentation and Outlook: Recording Seeding Parameters, Group Discussion, Q&A and Preparation for Day 2 Discussion & Q&A

Day 2: Exploring 3D Cell Models: Staining, Imaging & Interpretation

Time Activity Format
9:00 – 9:20 Welcome, Recap of Day 1, Lab Planning & Questions Q&A
9:20 – 10:00 Basics of Fluorescence Microscopy Lecture
10:00 – 10:45 Inspection of Participants’ Own 3D Cultures Laboratory
10:45 – 11:00 Coffee Break
11:00 – 12:30 Live/Dead Staining of Trainer-Prepared 72-Hour Spheroids: HepG2-GFP + PI + Hoechst or Alternative CellTracker Green + PI + Hoechst Laboratory
12:30 – 13:30 Lunch Break
13:30 – 15:30 Fluorescence Imaging (Depending on Group Size: Split Groups and Parallel Theory Sessions) Laboratory
15:30 – 16:30 3D Readouts and Assay Options; Group Discussion: Observations, Common Artefacts and Troubleshooting of 3D Cultures Lecture & Discussion

Day 3: From Imaging to Insight: Analysis & Scientific Communication

Time Activity Format
9:00 – 9:20 Welcome & Open Questions Q&A
9:20 – 10:30 Brightfield Imaging & Staining Laboratory
10:30 – 10:45 Coffee Break
10:45 – 12:30 Fluorescence Imaging (Depending on Duration and Sample Availability) Laboratory
12:30 – 13:30 Lunch Break
13:30 – 14:30 Preparation of Short Group Presentations (Optional Additional Fluorescence Imaging) Theory
14:30 – 15:30 Group Presentations & Scientific Discussion Theory
15:30 – 16:00 Final Wrap-Up, Feedback Session & Certificate Information Q&A

What Participants Will Gain

  • Hands-on experience with multiple 3D culture platforms
  • Practical laboratory skills in spheroid generation and maintenance
  • Introduction to fluorescence and confocal microscopy
  • Experience with live/dead staining workflows
  • Experimental planning and troubleshooting strategies
  • Scientific documentation and data interpretation skills
  • Exposure to modern applications in cancer biology, drug screening, organoids, and organ-on-chip technologies

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