title: Comparison of label-free and GFP multiphoton imaging of hair follicle-associated pluripotent (HAP) stem cells in mouse whiskers.
abstract: Hair-follicle-associated pluripotent (HAP) stem cells can differentiate into many cell types, including neurons and heart muscle cells, and have been shown to repair peripheral nerves and the spinal cord in mice. HAP stem cells can be obtained from each individual patient for regenerative medicine which overcomes problems with immune rejection. Previously, we have demonstrated that genetically-encoded protein markers such as GFP in transgenic mice can be used to visualize HAP stem cells in vivo by multiphoton tomography. Detection and visualization of stem cells in vivo without exogenous labels such as GFP would be important for human application. In the present report, we demonstrate label-free visualization of hair follicle stem cells in mouse whiskers by multiphoton tomography due to the intrinsic fluorophores such as NAD(P)H/flavins. We compared multiphoton tomography of GFP-labeled HAP stem cells and unlabeled stem cells in isolated mouse whiskers. We show that observation of HAP stem cells by label-free multiphoton tomography is comparable to detection using GFP-labeled stem cells. The results described here have important implications for detection and isolation of human HAP stem cells for regenerative medicine.
authors.affiliation: a Department of Biophotonics and Laser Technology ; Saarland University ; Saarbruecken , Germany.|b AntiCancer Inc. ; San Diego , CA USA.|c Department of Surgery ; University of California San Diego ; San Diego , CA USA.
authors.forename: Aisada
authors.initials: A
authors.lastname: Uchugonova
authors.affiliation: b AntiCancer Inc. ; San Diego , CA USA.|c Department of Surgery ; University of California San Diego ; San Diego , CA USA.
authors.forename: Wenluo
authors.initials: W
authors.lastname: Cao
authors.affiliation: b AntiCancer Inc. ; San Diego , CA USA.|c Department of Surgery ; University of California San Diego ; San Diego , CA USA.
authors.forename: Robert M
authors.initials: RM
authors.lastname: Hoffman
authors.affiliation: a Department of Biophotonics and Laser Technology ; Saarland University ; Saarbruecken , Germany.|d JenLab GmbH ; Jena , Germany.
authors.forename: Karsten
authors.initials: K
authors.lastname: Koenig
mesh_terms: D000818:Animals; D019070:Cell Lineage; D058948:Cell Tracking / Q000379:methods*; D017930:Genes, Reporter; D049452:Green Fluorescent Proteins / Q000096:biosynthesis* / Q000235:genetics; D018859:Hair Follicle / Q000166:cytology / Q000378:metabolism*; D008807:Mice, Inbred BALB C; D008822:Mice, Transgenic; D036641:Microscopy, Fluorescence, Multiphoton; D064231:Nestin / Q000235:genetics; D039904:Pluripotent Stem Cells / Q000378:metabolism*; D011401:Promoter Regions, Genetic; D014738:Vibrissae / Q000166:cytology / Q000378:metabolism*
chemical_list: C577485:Nes protein, mouse; D064231:Nestin; D049452:Green Fluorescent Proteins
keywords: GFP; hair follicles; multiphoton imaging; nestin; non-invasive imaging; pluripotent; skin; stem cells; tomography