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Improvement of Combined FISH and Immunofluorescence to Trace the Fate of Somatic Stem Cells after Transplantation

Chiara Donadoni, Stefania Corti, Federica Locatelli, Dimitra Papadimitriou, Michela Guglieri, Sandra Strazzer, Patrizia Bossolasco, Sabrina Salani and Giacomo P. Comi

Centro Dino Ferrari, Dipartimento di Scienze Neurologiche, Università degli Studi di Milano, IRCCS Ospedale Maggiore Policlinico, Milano, Italy (CD,SC,FL,DP,MG,SS,GPC); Center of Excellence on Neurodegenerative Diseases, Milano, Italy (SC,GPC); IRCCS Eugenio Medea, Bosisio Parini, Lecco, Italy (SS); and Fondazione Matarelli, Milano, Italy (PB)

Correspondence to: Prof. Giacomo P. Comi, Dipartimento di Scienze Neurologiche, Università di Milano, Padiglione Ponti, Ospedale Policlinico, Via Francesco Sforza 35, 20122 Milan, Italy. E-mail: giacomo.comi{at}unimi.it

Fluorescence in situ hybridization (FISH) combined with immunohistochemistry of tissue-specific markers provides a reliable method for characterizing the fate of somatic stem cells in transplantation experiments. Furthermore, the association between FISH and fluorescent gene reporter detection can unravel cell fusion phenomena, which could account for apparent transdifferentiation events. However, despite the widespread use of these techniques, they still require labor-extensive protocol adjustments to achieve correct and satisfactory simultaneous signal detection. In the present paper, we describe an improvement of simultaneous FISH and immunofluorescence detection. We applied this protocol to the identification of transplanted human and mouse hematopoietic stem cells in murine brain and muscle. This technique provides unique opportunities for following the path taken by transplanted cells and their differentiation into mature cell types.

(J Histochem Cytochem 52:1333–1339, 2004)

Key Words: fluorescence in situ hybridization • green fluorescent protein • stem cell • transplantation

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