Combined methods to evaluate human cells in muscle xenografts.
Journal
PloS one
ISSN: 1932-6203
Titre abrégé: PLoS One
Pays: United States
ID NLM: 101285081
Informations de publication
Date de publication:
2019
2019
Historique:
received:
15
01
2019
accepted:
19
04
2019
entrez:
4
5
2019
pubmed:
3
5
2019
medline:
14
1
2020
Statut:
epublish
Résumé
Xenotransplantation of human cells into immunodeficient mouse models is a very powerful tool and an essential step for the pre-clinical evaluation of therapeutic cell- and gene- based strategies. Here we describe an optimized protocol combining immunofluorescence and real-time quantitative PCR to both quantify and visualize the fate and localization of human myogenic cells after injection in regenerating muscles of immunodeficient mice. Whereas real-time quantitative PCR-based method provides an accurate quantification of human cells, it does not document their specific localization. The addition of an immunofluorescence approach using human-specific antibodies recognizing engrafted human cells gives information on the localization of the human cells within the host muscle fibres, in the stem cell niche or in the interstitial space. These two combined approaches offer an accurate evaluation of human engraftment including cell number and localization and should provide a gold standard to compare results obtained either using different types of human stem cells or comparing healthy and pathological muscle stem cells between different research laboratories worldwide.
Identifiants
pubmed: 31048846
doi: 10.1371/journal.pone.0211522
pii: PONE-D-19-01241
pmc: PMC6497248
doi:
Substances chimiques
DNA-Binding Proteins
0
Il2rb protein, mouse
0
Interleukin-2 Receptor beta Subunit
0
Rag2 protein, mouse
0
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
e0211522Déclaration de conflit d'intérêts
The authors have declared that no competing interests exist.
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