Hello, I’m Gherardo Baudo, a postdoctoral fellow with a Ph.D. in Nanomedicine. I specialize in drug delivery, particularly focusing on biomimetic lipid nanoparticles that encapsulate mRNA and drugs. My research spans diverse medical realms, including osteosarcoma, traumatic brain injury, and neurodegenerative diseases. Recognized as an expert in mitochondria transplantation, I employ this innovative strategy to address various cardiovascular conditions such as pulmonary arterial hypertension (PAH), atherosclerosis, and idiopathic pulmonary fibrosis (IPF). Currently, my research is dedicated to advancing our understanding of Alzheimer’s disease and exploring novel approaches for its treatment. With a comprehensive background in nanomedicine and a proven track record in translational research, I am committed to making significant contributions to the field of medical science.

University of Florence

Mechanical Engineering BE '17, University of Florence

Polytechnic Institute of Turin

Biomedical Engineering MS '20, Polytechnic Institute of Turin

PhD obtained from University Chinese Academy of Science November 2023

Pulmonary And Respiratory Medicine

Biomaterials

Molecular Biology

Oncology

Surgery

Developmental Neuroscience

Immunology

Biomedical Engineering

Physiology

Clinical Psychology

Pathology And Forensic Medicine

Urology

Cellular And Molecular Neuroscience

Ecology

Psychiatry And Mental Health

Public Health, Environmental And Occupational Health

Epidemiology

Periodontics

Biophysics

Endocrinology, Diabetes And Metabolism

Infectious Diseases

Pharmacology

Experimental Methods

Gherardo Baudo's Org

Standard Operating Procedure (SOP) for Transwell Migration Assay

DisclaimerThis SOP provides a general guideline for conducting cell migration assays using transwell chambers. It is crucial to note that specific parameters, including seeding density, incubation times, and concentrations of chemoattractants or inhibitors, may vary depending on the cell line and experimental objectives. Researchers are advised to optimize these parameters for their specific cell lines and experimental designs before proceeding.BackgroundCell migration is a process that plays a crucial role in various physiological and pathological conditions, including wound healing, immune response, and cancer metastasis. Understanding cell migration mechanisms can provide insights into the development and progression of diseases, as well as contribute to the development of novel therapeutic strategies. Migration assays, such as the transwell migration assay, are crucial in studying the motility of cells in vitro. These assays allow researchers to quantify the movement of cells through a semipermeable membrane towards a chemoattractant, mimicking the migration of cells in vivo. By manipulating the conditions under which these cells migrate, including the use of inducers or inhibitors that can either promote or inhibit migration, scientists can uncover the molecular pathways involved in cell movement and identify potential targets for therapy. ObjectiveTo measure the migratory response of cells to a chemoattractant or the effect of a migration inhibitor or inducer using a Transwell migration assay. Materials24-well plates8 µm Transwell® with 8.0 µm pore polycarbonate membrane insert (e.g., Greiner Bio-One)Cell culture medium with low serum concentration (e.g., 0.5% FBS)Chemoattractant of choiceMigration inhibitor or inducer (if applicable)4% Paraformaldehyde (PFA)0.1% Crystal violet solution90% Acetic acidCotton swabsMicroscopeMicroplate reader capable of reading absorbance at 590 nm Procedure Chemoattractant Addition: Fill the lower compartment of the well with 600 µL of medium containing the chemoattractant (inducer). If your chemoattractant is an inhibitor, you will resuspend it with the medium and the cells to evaluate whether the inhibitor affects the ability of the cells to move through the membrane (see steps 2 and 3).Preparation of Cell Suspensions: Resuspend cells in a low serum medium (e.g., 0.5% FBS) to a suitable concentration (e.g., 1×10^5 cells/mL).Seeding of Cells: Add 200 µL of the cell suspension to the upper compartment of the insert placed in a 24-well plate.Incubation: Incubate the plates for 24 hours (or appropriate time based on cell type and experimental needs) at 37 °C in a 5% CO2 atmosphere to allow cell migration through the pores.Fixation and Staining: After incubation, carefully remove the non-migrated cells from the upper surface of the membrane with a cotton swab. Fix migrated cells on the lower surface with 4% PFA for 15 min at room temperature. Wash twice with 1xPBS and then stain with 0.1% crystal violet for 30 min.Washing: Wash the membranes gently with 1xPBS to remove excess stain.Microscopic Imaging: After washing, take images of the stained migrated cells under the microscope for documentation and qualitative analysis.Dye Elution and Quantification: Elute the dye from the migrated cells with 90% acetic acid. Measure the absorbance of the eluted dye solution at 590 nm using a microplate reader. The absorbance is directly proportional to the number of migrated cells. Data AnalysisAnalyze the data by comparing the absorbance values from wells with chemoattractant alone to those with chemoattractant and migration inducer or inhibitor. Use control wells with cells and low serum medium without chemoattractant for background correction.  The schematic illustration included in this SOP is a visual representation of the transwell migration assay process and was generated using BioRender. This diagram is intended to provide a clear and concise overview of the experimental setup and procedural steps.

Standard Operating Procedure (SOP) for Transwell Migration Assay

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