Cell Culture Course

Lesson 1 • Cell Biology Essentials for Culture

Covers cell structure, organelle function, and growth requirements relevant to in vitro work. Provides the biological rationale behind every culture decision made later.

Lesson 2 • Aseptic Technique Principles

Introduces contamination sources, airflow dynamics, and behavioural practices that prevent microbial ingress. Forms the behavioural foundation for every hands-on procedure in the course.

Lesson 3 • History and Scope of Cell Culture

Traces development from early tissue explants to modern 3D systems. Contextualises why standardised practices emerged and how they shape current workflows.

Lesson 1 • Media Preparation and Quality Control

Covers sterile filtration, pH adjustment, aliquoting, and shelf-life testing of prepared media. Connects QC practices to downstream experimental reliability.

Lesson 2 • Antibiotics and Antifungals in Media

Evaluates appropriate use, concentrations, and risks of antimicrobial agents in culture. Addresses dependency risks and when antibiotic-free culture is preferred.

Lesson 3 • Specialised and Custom Media Formulations

Introduces organ-specific, 3D culture, and chemically defined media for advanced applications. Prepares students to adapt formulations to non-standard cell types.

Lesson 4 • Serum and Serum-Free Supplements

Compares serum-containing and defined serum-free formulations, including growth factors and hormones. Guides selection based on experimental reproducibility and ethical considerations.

Lesson 1 • Incubator Types and Operating Principles

Compares CO2, tri-gas, and hypoxic incubators and their internal control mechanisms. Aligns incubator selection to the oxygen and CO2 demands of specific cell types.

Lesson 2 • Equipment Maintenance and Calibration Logs

Establishes preventive maintenance schedules, calibration documentation, and equipment failure response. Links proper records to regulatory compliance and data integrity.

Lesson 3 • Centrifuge and Microscope Use

Teaches proper centrifuge balancing, speed selection, and rotor care alongside basic microscopy for culture assessment. Integrates equipment use into daily culture workflow.

Lesson 4 • Gas Phase and pH Regulation

Explains CO2 and bicarbonate buffering equilibrium and its effect on media pH stability. Teaches calibration and monitoring routines to maintain physiological pH.

Lesson 1 • Cell Line Sourcing and Authentication

Covers acquisition from repositories, STR profiling, and mycoplasma testing for identity assurance. Prevents use of misidentified or cross-contaminated lines in experiments.

Lesson 2 • Primary Cell Isolation Techniques

Introduces enzymatic digestion, mechanical dissociation, and density gradient separation for tissue-derived cells. Connects isolation method choice to downstream cell viability and purity.

Lesson 3 • Routine Subculture and Passaging

Teaches trypsinisation, cell counting, seeding density calculation, and passage recording. Establishes consistent passaging habits that preserve cell phenotype over time.

Lesson 4 • Cryopreservation and Cell Banking

Details cryoprotectant selection, controlled-rate freezing, and liquid nitrogen storage protocols. Ensures students can build and recover working and master cell banks.

Lesson 1 • Mycoplasma Detection and Elimination

Covers PCR, DAPI staining, and commercial kit methods for mycoplasma screening. Addresses elimination strategies and quarantine protocols for positive cultures.

Lesson 2 • Chemical and Cross-Contamination Sources

Identifies endotoxins, plasticizer leaching, and reagent carryover as non-microbial contamination risks. Connects contamination source identification to corrective material or process changes.

Lesson 3 • Microbial Contamination Recognition

Identifies visual and microscopic signs of bacterial, fungal, and yeast contamination in cultures. Trains rapid recognition to minimize experimental loss and cross-contamination spread.

Lesson 4 • Decontamination and Disinfection Methods

Compares autoclaving, chemical disinfectants, UV irradiation, and dry heat for surface and material sterilization. Guides selection based on material compatibility and efficacy data.

Lesson 1 • Viability Assay Principles and Methods

Compares trypan blue exclusion, metabolic assays, and membrane integrity dyes for viability measurement. Links assay choice to sensitivity requirements and throughput needs.

Lesson 2 • Cell Identity and Phenotype Verification

Uses immunofluorescence, flow cytometry markers, and functional assays to confirm cell identity. Prevents phenotypic drift from invalidating experimental conclusions.

Lesson 3 • Proliferation and Growth Curve Analysis

Covers doubling time calculation, growth curve construction, and population doubling level tracking. Provides quantitative benchmarks for detecting phenotypic drift across passages.

Lesson 4 • Apoptosis and Cell Death Assays

Introduces annexin V, caspase activity, and TUNEL assays to distinguish apoptosis from necrosis. Connects death pathway identification to experimental interpretation and troubleshooting.

Lesson 1 • Viral Vector Transduction

Covers lentiviral, adenoviral, and AAV vector production, titration, and transduction protocols. Addresses biosafety requirements and tropism considerations for vector selection.

Lesson 2 • Physical Transfection Methods

Introduces electroporation, nucleofection, and microinjection as physical delivery approaches. Addresses parameter optimisation for hard-to-transfect primary and suspension cells.

Lesson 3 • CRISPR-Cas9 Genome Editing in Culture

Applies CRISPR-Cas9 delivery, guide RNA design, and editing efficiency validation in cultured cells. Connects editing confirmation workflows to downstream functional studies.

Lesson 4 • Principles of Gene Delivery

Explains barriers to nucleic acid entry and how physical, chemical, and viral methods overcome them. Frames delivery mechanism understanding as the basis for method selection.

Lesson 1 • Suspension and Bioreactor Culture

Covers stirred-tank, wave, and hollow-fiber bioreactor configurations for suspension and adherent cells. Addresses agitation, aeration, and feeding strategies for high-density culture.

Lesson 2 • Three-Dimensional Culture Models

Compares spheroids, organoids, and scaffold-based systems for physiological relevance over 2D monolayers. Guides model selection based on research question and downstream assay compatibility.

Lesson 3 • Good Manufacturing Practice in Cell Culture

Introduces GMP documentation, change control, and batch record requirements for regulated cell culture. Prepares students to operate within quality management systems for cell-based products.

Lesson 4 • Microfluidic and Organ-on-a-Chip Systems

Introduces microfluidic device design, flow parameters, and multi-organ co-culture configurations. Positions these platforms as bridges between cell culture and in vivo physiology.