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.

Broggi, A., Cigni, C., Zanoni, I., & Granucci, F. (2016). Preparation of single-cell suspensions for cytofluorimetric analysis from different mouse skin regions. JOURNAL OF VISUALIZED EXPERIMENTS, 2016(110) [10.3791/52589].

Preparation of single-cell suspensions for cytofluorimetric analysis from different mouse skin regions

BROGGI, ACHILLE
Primo
;
CIGNI, CLARA
Secondo
;
ZANONI, IVAN
Penultimo
;
GRANUCCI, FRANCESCA
Ultimo
2016

Abstract

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.
Si
Articolo in rivista - Articolo scientifico
Scientifica
Collagenase; Dermis; Flow cytometry; Immune system; Issue 110; Leukocytes; Medicine; Single-cell preparation; Skin;
Collagenase; Dermis; Flow cytometry; Immune system; Issue 110; Leukocytes; Medicine; Single-cell preparation; Skin; Biochemistry, Genetics and Molecular Biology (all); Chemical Engineering (all); Immunology and Microbiology (all); Neuroscience (all)
English
Broggi, A., Cigni, C., Zanoni, I., & Granucci, F. (2016). Preparation of single-cell suspensions for cytofluorimetric analysis from different mouse skin regions. JOURNAL OF VISUALIZED EXPERIMENTS, 2016(110) [10.3791/52589].
Broggi, A; Cigni, C; Zanoni, I; Granucci, F
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Utilizza questo identificativo per citare o creare un link a questo documento: http://hdl.handle.net/10281/134599
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