Isolation of multipotent stem cells from mouse adipose tissue
Isolation of multipotent stem cells from mouse adipose tissue
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DOI：
10.1016/j.jdermsci.2007.05.015
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发表时间：
2007-10-01
影响因子：
4.6
通讯作者：
Matsunaga, Kayoko
中科院分区：
文献类型：
--
作者：
Yamamoto, Naoki;Akamatsu, Hirohiko;Matsunaga, Kayoko
背景资料：胚胎干细胞（ES）、骨髓、脂肪组织或其他遗传修饰的干细胞正被广泛用于再生医学领域的基础研究。然而，没有特异性的表面抗原可以用作多能干细胞的标记。目的：方法：采用抗CD 90、抗CD 105和抗Sca-1抗体作为间充质干细胞（MSC）的标记物，抗p75 NTR抗体作为上皮干细胞和MSC的标记物，对小鼠皮下脂肪组织进行免疫组化染色。随后，通过流式细胞术（荧光激活细胞分选：FACS）检查脂肪组织来源的基质血管组分培养细胞（ADSVF细胞）中细胞表面标志物的表达。最后，将p75 NTR阳性的ADSVF细胞进行分选和培养，诱导其分化为脂肪细胞、成骨细胞、软骨细胞、平滑肌细胞和神经元细胞。其中p75 NTR阳性细胞分化为脂肪细胞、成骨细胞、软骨细胞、平滑肌细胞和神经细胞。结论：p75 NTR可作为分离脂肪组织源性干细胞（ASCs）的有效标志物。(c)2007年日本皮肤病研究学会。由Elsevier爱尔兰有限公司出版。保留所有权利。
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 muttipotent 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, osteobtasts, 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, osteobtasts, chondrocytes, smooth muscle cells and neuronal cells. The rate of differentiation into adipocytes, osteobtasts 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).(c) 2007 Japanese Society for Investigative Dermatology. Published by Elsevier Ireland Ltd. All rights reserved.