Isolation of multipotent stem cells from mouse adipose tissue

Naoki Yamamoto, Hirohiko Akamatsu, Seiji Hasegawa, Takaaki Yamada, Satoru Nakata, Mahito Ohkuma, Ei Ichi Miyachi, Tohru Marunouchi, Kayoko Matsunaga

Research output: Contribution to journalArticle

122 Citations (Scopus)

Abstract

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).

Original languageEnglish
Pages (from-to)43-52
Number of pages10
JournalJournal of Dermatological Science
Volume48
Issue number1
DOIs
Publication statusPublished - 01-10-2007

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Multipotent Stem Cells
Nerve Growth Factor Receptor
Stem cells
Adipose Tissue
Tissue
Cells
Osteoblasts
Adipocytes
Stem Cells
Chondrocytes
Mesenchymal Stromal Cells
Smooth Muscle Myocytes
Blood Vessels
Cell culture
Flow Cytometry
Cell Culture Techniques
Muscle
Regenerative Medicine
Subcutaneous Fat
Surface Antigens

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Molecular Biology
  • Dermatology

Cite this

Yamamoto, N., Akamatsu, H., Hasegawa, S., Yamada, T., Nakata, S., Ohkuma, M., ... Matsunaga, K. (2007). Isolation of multipotent stem cells from mouse adipose tissue. Journal of Dermatological Science, 48(1), 43-52. https://doi.org/10.1016/j.jdermsci.2007.05.015
Yamamoto, Naoki ; Akamatsu, Hirohiko ; Hasegawa, Seiji ; Yamada, Takaaki ; Nakata, Satoru ; Ohkuma, Mahito ; Miyachi, Ei Ichi ; Marunouchi, Tohru ; Matsunaga, Kayoko. / Isolation of multipotent stem cells from mouse adipose tissue. In: Journal of Dermatological Science. 2007 ; Vol. 48, No. 1. pp. 43-52.
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Yamamoto, N, Akamatsu, H, Hasegawa, S, Yamada, T, Nakata, S, Ohkuma, M, Miyachi, EI, Marunouchi, T & Matsunaga, K 2007, 'Isolation of multipotent stem cells from mouse adipose tissue', Journal of Dermatological Science, vol. 48, no. 1, pp. 43-52. https://doi.org/10.1016/j.jdermsci.2007.05.015

Isolation of multipotent stem cells from mouse adipose tissue. / Yamamoto, Naoki; Akamatsu, Hirohiko; Hasegawa, Seiji; Yamada, Takaaki; Nakata, Satoru; Ohkuma, Mahito; Miyachi, Ei Ichi; Marunouchi, Tohru; Matsunaga, Kayoko.

In: Journal of Dermatological Science, Vol. 48, No. 1, 01.10.2007, p. 43-52.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Isolation of multipotent stem cells from mouse adipose tissue

AU - Yamamoto, Naoki

AU - Akamatsu, Hirohiko

AU - Hasegawa, Seiji

AU - Yamada, Takaaki

AU - Nakata, Satoru

AU - Ohkuma, Mahito

AU - Miyachi, Ei Ichi

AU - Marunouchi, Tohru

AU - Matsunaga, Kayoko

PY - 2007/10/1

Y1 - 2007/10/1

N2 - 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).

AB - 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).

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