title: Flow cytometric and immunohistochemical detection of in vivo BrdU-labeled cells in mouse fat depots.
abstract: This study has determined the natural frequency and localization of progenitor/stem cells within fat depots in situ based on their ability to retain DNA nucleotide label (BrdU). Neonate and mature male C57BL6/J mice were injected intraperitoneally with BrdU- and label-retaining cells (LRC) were quantified in fat depots by immunohistochemical, immunofluorescent, and flow cytometric methods. In neonates, LRC constituted 27% of the cells in inguinal fat (iWAT) and 65% in interscapular brown fat (BAT) after Day 10 and 26% of the cells in epididymal fat (eWAT) after Day 28. After 52 days, the LRC accounted for 0.72% of iWAT, 0.53% of eWAT and 1.05% of BAT, respectively. The BrdU-labeled cells localized to two areas: single cells distributed among adipocytes or those adjacent to the blood vessels wall. In mature C57BL6/J mice, flow cytometric analysis determined that a majority of the LRC were also positive for stem cell antigen-1 (Sca-1).
authors.affiliation: Pennington Biomedical Research Center, Louisiana State University System, 6400 Perkins Rd., Baton Rouge, LA 70808, USA.
authors.forename: Jaroslaw
authors.initials: J
authors.lastname: Staszkiewicz
authors.forename: Jeffrey
authors.initials: J
authors.lastname: Gimble
authors.forename: Courtney
authors.initials: C
authors.lastname: Cain
authors.forename: Marilyn
authors.initials: M
authors.lastname: Dietrich
authors.forename: David
authors.initials: D
authors.lastname: Burk
authors.forename: Heather
authors.initials: H
authors.lastname: Kirk-Ballard
authors.forename: Barbara
authors.initials: B
authors.lastname: Gawronska-Kozak
mesh_terms: D002001:Adipose Tissue, Brown / Q000166:cytology* / Q000378:metabolism; D052436:Adipose Tissue, White / Q000166:cytology* / Q000378:metabolism; D000818:Animals; D000950:Antigens, Ly / Q000032:analysis / Q000378:metabolism; D001973:Bromodeoxyuridine / Q000032:analysis* / Q000378:metabolism; D004822:Epididymis / Q000166:cytology* / Q000378:metabolism; D005434:Flow Cytometry; D007150:Immunohistochemistry; D008297:Male; D008565:Membrane Proteins / Q000032:analysis / Q000378:metabolism; D051379:Mice; D008810:Mice, Inbred C57BL; D013194:Staining and Labeling; D013234:Stem Cells / Q000166:cytology*
chemical_list: D000950:Antigens, Ly; C074650:Ly6a protein, mouse; D008565:Membrane Proteins; D001973:Bromodeoxyuridine