Organotypic culture of human bone marrow adipose tissue
Organotypic culture of human bone marrow adipose tissue
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DOI：
10.1111/j.1440-1827.2010.02511.x
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发表时间：
2010-04
影响因子：
2.2
通讯作者：
K. Uchihashi;S. Aoki;M. Shigematsu;Noriyuki Kamochi;Emiko Sonoda;H. Soejima;K. Fukudome;H. Sugihara;T. Hotokebuchi;S. Toda
中科院分区：
文献类型：
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作者：
K. Uchihashi;S. Aoki;M. Shigematsu;Noriyuki Kamochi;Emiko Sonoda;H. Soejima;K. Fukudome;H. Sugihara;T. Hotokebuchi;S. Toda
骨髓脂肪组织（BMAT）在骨髓中的确切作用尚不清楚。因此，本研究的目的是描述一种利用BMAT片段的三维胶原凝胶培养、免疫组织化学、ELISA和实时逆转录聚合酶链反应来研究BMAT的新方法。成熟脂肪细胞和CD45+白细胞保留50 ~ 3周。骨髓基质细胞（BMSC）包括少量脂质富集的前脂肪细胞和CD44+/CD105+间充质干细胞（MSC）样细胞，由BMAT发育而来。地塞米松（10 μ mol/L）显著增加了前脂肪细胞的数量，而胰岛素（20 μ mol/ mL）没有显著增加。地塞米松和胰岛素也能促进BMAT中瘦素的产生和基因表达。在所有培养条件下，BMAT的脂联素产量均<0.8 ng/mL。地塞米松促进脂联素基因表达，胰岛素抑制脂联素基因表达。这一发现表明地塞米松，而不是胰岛素，可能是BMAT的一个强大的脂肪生成因素，其中脂联素蛋白分泌通常非常低，BMAT可能表现出与内脏和皮下脂肪组织不同的表型。BMAT与成骨细胞的相互作用也被检测，发现成骨细胞抑制BMSC的发育并减少瘦素的产生，而BMAT抑制成骨细胞的生长和分化。该方法对BMAT生物学的研究具有重要意义。
The precise role of bone marrow adipose tissue (BMAT) in the marrow remains unknown. The purpose of the present study was therefore to describe a novel method for studying BMAT using 3‐D collagen gel culture of BMAT fragments, immunohistochemistry, ELISA and real‐time reverse transcription–polymerase chain reaction. Mature adipocytes and CD45+ leukocytes were retained for >3 weeks. Bone marrow stromal cells (BMSC) including a small number of lipid‐laden preadipocytes and CD44+/CD105+ mesenchymal stem cell (MSC)‐like cells, developed from BMAT. Dexamethasone (10 µmol/L), but not insulin (20 mU/mL), significantly increased the number of preadipocytes. Dexamethasone and insulin also promoted leptin production and gene expression in BMAT. Adiponectin production by BMAT was <0.8 ng/mL under all culture conditions. Dexamethasone promoted adiponectin gene expression, while insulin inhibited it. This finding suggests that dexamethasone, but not insulin, may serve as a powerful adipogenic factor for BMAT, in which adiponectin protein secretion is normally very low, and that BMAT may exhibit a different phenotype from that of the visceral and subcutaneous adipose tissues. BMAT–osteoblast interactions were also examined, and it was found that osteoblasts inhibited the development of BMSC and reduced leptin production, while BMAT inhibited the growth and differentiation of osteoblasts. The present novel method proved to be useful for the study of BMAT biology.