Preparation of Single-cell Suspensions for Cytofluorimetric Analysis from Different Mouse Skin Regions

Autor: Clara Cigni, Francesca Granucci, Ivan Zanoni, Achille Broggi
Přispěvatelé: Broggi, A, Cigni, C, Zanoni, I, Granucci, F
Rok vydání: 2016
Předmět:
CD4-Positive T-Lymphocytes
0301 basic medicine
Pathology
Immunology and Microbiology (all)
General Chemical Engineering
Collagenase
Cell
Cell Separation
CD8-Positive T-Lymphocytes
Mice
Skin Physiological Phenomena
Chemical Engineering (all)
Skin
Single-cell preparation
integumentary system
medicine.diagnostic_test
General Neuroscience
Flow Cytometry
Cell biology
Matrix Metalloproteinase 8
medicine.anatomical_structure
Dermi
Medicine
medicine.medical_specialty
Thermolysin
Biology
General Biochemistry
Genetics and Molecular Biology
Flow cytometry
Issue 110
03 medical and health sciences
Immune system
Suspensions
Antigen
Dermis
medicine
Animals
Collagenases
Viability assay
Biochemistry
Genetics and Molecular Biology (all)
Neuroscience (all)
General Immunology and Microbiology
Epidermis (botany)
Macrophages
Dendritic Cells
Leukocyte
030104 developmental biology
Epidermal Cells
Biomarkers
Zdroj: Journal of Visualized Experiments.
ISSN: 1940-087X
Popis: The skin is a barrier organ that interacts with the external environment. Being continuously exposed to potential microbial invasion, the dermis and epidermis home a variety of immune cells in both homeostatic and inflammatory conditions. Tools to obtain skin cell release for cytofluorimetric analyses are, therefore, very useful in order to study the complex network of immune cells residing in the skin and their response to microbial stimuli. Here, we describe an efficient methodology for the digestion of mouse skin to rapidly and efficiently obtain single-cell suspensions. This protocol allows maintenance of maximum cell viability without compromising surface antigen expression. We also describe how to take and digest skin samples from different anatomical locations, such as the ear, trunk, tail, and footpad. The obtained suspensions are then stained and analyzed by flow cytometry to discriminate between different leukocyte populations.
Databáze: OpenAIRE
Externí odkaz: