Isolation of multipotent stem cells from mouse adipose tissue.
BACKGROUND
Embryonic stem (ES) cells, bone marrow, adipose tissue or other genetically modified stem cells are being widely used in basic research in the field of regenerative medicine. However, there is no specific surface antigen that can be used as a marker of multipotent stem cells.


OBJECTIVE
We tried to isolate and collect putative multipotent stem cells from mouse subcutaneous adipose tissue using the p75 neurotrophin receptor (p75NTR) as a marker.


METHODS
Adipose tissue was processed for immunostaining using antibodies anti-CD90, anti-CD105 and anti-Sca-1 as general mesenchymal stem cell (MSC) markers, and anti-p75NTR, an epithelial stem cell and MSC marker. Subsequently, the expression of cell surface markers in adipose tissue-derived stromal vascular fraction culture cells (ADSVF cells) was examined by flow cytometry (fluorescence-activated cell sorting: FACS). Finally, ADSVF cells positive for p75NTR were sorted and cultured to induce their differentiation into adipocytes, osteoblasts, chondrocytes, smooth muscle cells and neuronal cells.


RESULTS
Cells positive for several of these markers were found in the deep layers of adipose tissue. Among them, those positive for p75NTR differentiated into adipocytes, osteoblasts, chondrocytes, smooth muscle cells and neuronal cells. The rate of differentiation into adipocytes, osteoblasts and neuronal cells was higher for p75NTR-positive cells than for p75NTR-negative cells.


CONCLUSIONS
p75NTR proved to be a useful marker to isolate adipose tissue-derived stem cells (ASCs).
Yamamoto
Naoki
N
Laboratory of Molecular Biology & Histochemistry, Fujita Health University Joint Research Laboratory, 1-98 Toyoake, Aichi 470-1192, Japan. naokiy@fujita-hu.ac.jp
Akamatsu
Hirohiko
H
Hasegawa
Seiji
S
Yamada
Takaaki
T
Nakata
Satoru
S
Ohkuma
Mahito
M
Miyachi
Ei-Ichi
E
Marunouchi
Tohru
T
Matsunaga
Kayoko
K
D017667:Adipocytes / Q000166:cytology / Q000378:metabolism; D000273:Adipose Tissue / Q000166:cytology* / Q000378:metabolism; D000818:Animals; D002469:Cell Separation / Q000379:methods*; D002478:Cells, Cultured; D005434:Flow Cytometry / Q000379:methods; D008297:Male; D051379:Mice; D008813:Mice, Inbred ICR; D039902:Multipotent Stem Cells / Q000166:cytology* / Q000378:metabolism; D010006:Osteoblasts / Q000166:cytology / Q000378:metabolism; D017475:Receptors, Nerve Growth Factor / Q000378:metabolism
D017475:Receptors, Nerve Growth Factor; C517634:Ngfr protein, mouse