Characterization of transplanted green fluorescent protein+ bone marrow cells into adipose tissue.
Following transplantation of green fluorescent protein (GFP)-labeled bone marrow (BM) into irradiated, wild-type Sprague-Dawley rats, propagated GFP(+) cells migrate to adipose tissue compartments. To determine the relationship between GFP(+) BM-derived cells and tissue-resident GFP(-) cells on the stem cell population of adipose tissue, we conducted detailed immunohistochemical analysis of chimeric whole fat compartments and subsequently isolated and characterized adipose-derived stem cells (ASCs) from GFP(+) BM chimeras. In immunohistochemistry, a large fraction of GFP(+) cells in adipose tissue were strongly positive for CD45 and smooth muscle actin and were evenly scattered around the adipocytes and blood vessels, whereas all CD45(+) cells within the blood vessels were GFP(+). A small fraction of GFP(+) cells with the mesenchymal marker CD90 also existed in the perivascular area. Flow cytometric and immunocytochemical analyses showed that cultured ASCs were CD45(-)/CD90(+)/CD29(+). There was a significant difference in both the cell number and phenotype of the GFP(+) ASCs in two different adipose compartments, the omental (abdominal) and the inguinal (subcutaneous) fat pads; a significantly higher number of GFP(-)/CD90(+) cells were isolated from the subcutaneous depot as compared with the abdominal depot. The in vitro adipogenic differentiation of the ASCs was achieved; however, all cells that had differentiated were GFP(-). Based on phenotypical analysis, GFP(+) cells in adipose tissue in this rat model appear to be of both hematopoietic and mesenchymal origin; however, infrequent isolation of GFP(+) ASCs and their lack of adipogenic differentiation suggest that the contribution of BM to ASC generation might be minor.
Tomiyama
Koji
K
Thomas E. Starzl Transplantation Institute, Department of Surgery, University of Pittsburgh, 200 Lothrop Street, BST 1655E, Pittsburgh, Pennsylvania 15261, USA.
Murase
Noriko
N
Stolz
Donna Beer
DB
Toyokawa
Hideyoshi
H
O'Donnell
Daniel R
DR
Smith
Darren M
DM
Dudas
Jason R
JR
Rubin
J Peter
JP
Marra
Kacey G
KG
D000273:Adipose Tissue / Q000166:cytology* / Q000378:metabolism*; D053687:Adult Stem Cells / Q000166:cytology / Q000378:metabolism; D000818:Animals; D030801:Animals, Genetically Modified; D015703:Antigens, CD / Q000378:metabolism; D001854:Bone Marrow Cells / Q000166:cytology* / Q000378:metabolism*; D016026:Bone Marrow Transplantation; D002454:Cell Differentiation; D049452:Green Fluorescent Proteins / Q000378:metabolism*; D006412:Hematopoietic Stem Cells / Q000166:cytology / Q000378:metabolism; D059630:Mesenchymal Stem Cells / Q000166:cytology / Q000378:metabolism; D046529:Microscopy, Electron, Transmission; D008856:Microscopy, Fluorescence; D010641:Phenotype; D011828:Radiation Chimera; D051381:Rats; D017207:Rats, Sprague-Dawley; D011994:Recombinant Proteins / Q000378:metabolism
D015703:Antigens, CD; D011994:Recombinant Proteins; D049452:Green Fluorescent Proteins