Flow cytometric and immunohistochemical detection of in vivo BrdU-labeled cells in mouse fat depots

Biochem Biophys Res Commun. 2009 Jan 16;378(3):539-44. doi: 10.1016/j.bbrc.2008.11.092. Epub 2008 Dec 3.

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).

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Adipose Tissue, Brown / cytology*
  • Adipose Tissue, Brown / metabolism
  • Adipose Tissue, White / cytology*
  • Adipose Tissue, White / metabolism
  • Animals
  • Antigens, Ly / analysis
  • Antigens, Ly / metabolism
  • Bromodeoxyuridine / analysis*
  • Bromodeoxyuridine / metabolism
  • Epididymis / cytology*
  • Epididymis / metabolism
  • Flow Cytometry
  • Immunohistochemistry
  • Male
  • Membrane Proteins / analysis
  • Membrane Proteins / metabolism
  • Mice
  • Mice, Inbred C57BL
  • Staining and Labeling
  • Stem Cells / cytology*

Substances

  • Antigens, Ly
  • Ly6a protein, mouse
  • Membrane Proteins
  • Bromodeoxyuridine