Isolation and Characterization of Multipotent Mesenchymal Stem Cells Adhering to Adipocytes in Canine Bone Marrow
Isolation and Characterization of Multipotent Mesenchymal Stem Cells Adhering to Adipocytes in Canine Bone Marrow
复制标题
DOI：
10.1089/scd.2016.0200
复制
发表时间：
2017-03-01
影响因子：
4
通讯作者：
Nishimura, Ryohei
中科院分区：
文献类型：
--
作者：
Lin, Hsing-Yi;Fujita, Naoki;Nishimura, Ryohei
天花板培养方法已被用于从脂肪组织中分离成熟脂肪细胞，其可以去分化为成纤维细胞，也称为去分化脂肪（DFAT）细胞，其自我更新并且是多能的，具有比脂肪组织来源的间充质干细胞高得多的同质性和集落形成效率。我们使用这种技术培养犬骨髓脂肪细胞，期望获得DFAT细胞。然而，与我们的预期相反，通过延时显微镜对天花板培养物的连续监测显示许多小细胞粘附于脂肪细胞，其迅速增殖成纤维细胞形态的细胞，并且没有从脂肪细胞的任何去分化。我们将这些细胞命名为骨髓周脂肪细胞（BM-PAC），并证明了BM-PAC与传统培养的犬骨髓间充质干细胞（BMMSCs）相比的多能特性。BM-PAC的克隆形成能力和增殖能力明显强于BMMSCs。体外三系分化实验表明BM-PAC具有上级的成脂、成骨和成软骨能力。流式细胞术分析显示，在细胞生长和分化中起重要作用的CD 73的表达在BM-PAC中显著高于BMMSCs。这些结果表明，犬BM-PAC的干细胞特性是上级的BMMSCs，这些间充质干细胞（MSC）似乎是一个可行的来源，为基于细胞的治疗在狗。
The ceiling culture method has been used to isolate mature adipocytes from adipose tissue that can be dedifferentiated into fibroblastic cells, also known as dedifferentiated fat (DFAT) cells that self-renew and are multipotent, with much higher homogeneity and colony-forming efficiency than those of adipose tissue-derived mesenchymal stem cells. We cultured adipocytes from canine bone marrow using this technique, with the expectation of obtaining DFAT cells. However, contrary to our expectations, continuous monitoring of ceiling cultures by time-lapse microscopy revealed many small cells adhering to adipocytes that proliferated rapidly into cells with a fibroblastic morphology and without any dedifferentiation from adipocytes. We named these cells bone marrow peri-adipocyte cells (BM-PACs) and demonstrated the multipotent properties of BM-PACs compared to that of conventionally cultured canine bone marrow mesenchymal stem cells (BMMSCs). BM-PACs showed significantly greater clonogenicity and proliferation ability than BMMSCs. An in vitro trilineage differentiation assay revealed that BM-PACs possess adipogenic, osteogenic, and chondrogenic capacities superior to those of BMMSCs. Flow cytometric analysis revealed that the expression of CD73, which plays an important role in cell growth and differentiation, was significantly higher in BM-PACs than in BMMSCs. These results indicate that canine BM-PACs have stem cell characteristics that are superior to those of BMMSCs, and that these mesenchymal stem cells (MSCs) appear to be a feasible source for cell-based therapies in dogs.