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, Giacomo P. Comi

Research output: Contribution to journalArticle

Abstract

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.

Original languageEnglish
Pages (from-to)1333-1339
Number of pages7
JournalJournal of Histochemistry and Cytochemistry
Volume52
Issue number10
DOIs
Publication statusPublished - Oct 2004

Fingerprint

Adult Stem Cells
Stem Cell Transplantation
Fluorescence In Situ Hybridization
Fluorescent Antibody Technique
Forensic Anthropology
Cell Fusion
Hematopoietic Stem Cells
Reporter Genes
Cell Differentiation
Immunohistochemistry
Muscles
Brain

Keywords

  • Fluorescence in situ hybridization
  • Green fluorescent protein
  • Stem cell
  • Transplantation

ASJC Scopus subject areas

  • Anatomy
  • Cell Biology

Cite this

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abstract = "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.",
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T1 - Improvement of combined FISH and immunofluorescence to trace the fate of somatic stem cells after transplantation

AU - Donadoni, Chiara

AU - Corti, Stefania

AU - Locatelli, Federica

AU - Papadimitriou, Dimitra

AU - Guglieri, Michela

AU - Strazzer, Sandra

AU - Bossolasco, Patrizia

AU - Salani, Sabrina

AU - Comi, Giacomo P.

PY - 2004/10

Y1 - 2004/10

N2 - 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.

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KW - Fluorescence in situ hybridization

KW - Green fluorescent protein

KW - Stem cell

KW - Transplantation

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