Flow cytometric and immunohistochemical detection of in vivo BrdU-labeled cells in mouse fat depots.
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).
Staszkiewicz
Jaroslaw
J
Pennington Biomedical Research Center, Louisiana State University System, 6400 Perkins Rd., Baton Rouge, LA 70808, USA.
Gimble
Jeffrey
J
Cain
Courtney
C
Dietrich
Marilyn
M
Burk
David
D
Kirk-Ballard
Heather
H
Gawronska-Kozak
Barbara
B
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*
D000950:Antigens, Ly; C074650:Ly6a protein, mouse; D008565:Membrane Proteins; D001973:Bromodeoxyuridine