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Use of osmium tetroxide staining with microcomputerized tomography to visualize and quantify bone marrow adipose tissue in vivo.

Abstract
Adipocytes reside in discrete, well-defined depots throughout the body. In addition to mature adipocytes, white adipose tissue depots are composed of many cell types, including macrophages, endothelial cells, fibroblasts, and stromal cells, which together are referred to as the stromal vascular fraction (SVF). The SVF also contains adipocyte progenitors that give rise to mature adipocytes in those depots. Marrow adipose tissue (MAT) or marrow fat has long been known to be present in bone marrow (BM) but its origin, development, and function remain largely unknown. Clinically, increased MAT is associated with age, metabolic diseases, drug treatment, and marrow recovery in children receiving radiation and chemotherapy. In contrast to the other depots, MAT is unevenly distributed in the BM of long bones. Conventional quantitation relies on sectioning of the bone to overcome issues with distribution but is time-consuming, resource intensive, inconsistent between laboratories and may be unreliable as it may miss changes in MAT volume. Thus, the inability to quantitate MAT in a rapid, systematic, and reproducible manner has hampered a full understanding of its development and function. In this chapter, we describe a new technique that couples histochemical staining of lipid using osmium tetroxide with microcomputerized tomography to visualize and quantitate MAT within the medullary canal in three dimensions. Imaging of osmium staining provides a high-resolution map of existing and developing MAT in the BM. Because this method is simple, reproducible, and quantitative, we expect it will become a useful tool for the precise characterization of MAT.
AuthorsErica L Scheller, Nancy Troiano, Joshua N Vanhoutan, Mary A Bouxsein, Jackie A Fretz, Yougen Xi, Tracy Nelson, Griffin Katz, Ryan Berry, Christopher D Church, Casey R Doucette, Matthew S Rodeheffer, Ormond A Macdougald, Clifford J Rosen, Mark C Horowitz
JournalMethods in enzymology (Methods Enzymol) Vol. 537 Pg. 123-39 ( 2014) ISSN: 1557-7988 [Electronic] United States
PMID24480344 (Publication Type: Journal Article, Research Support, N.I.H., Extramural, Research Support, Non-U.S. Gov't)
Copyright© 2014 Elsevier Inc. All rights reserved.
Chemical References
  • Osmium Tetroxide
Topics
  • Adipogenesis (genetics)
  • Adipose Tissue, White (growth & development)
  • Bone Marrow (growth & development)
  • Cell Differentiation
  • Humans
  • Osmium Tetroxide (chemistry)
  • Staining and Labeling (methods)
  • Stromal Cells (cytology)
  • X-Ray Microtomography (methods)

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