Kassis I, Zangi L, Rivkin R, Levdansky L, Samuel S, Marx G, Gorodetsky R
Radiobiology and Biotechnology Laboratory, Sharett Institute of Oncology, Hadassah-Hebrew University Medical Center, Jerusalem, Israel.
Bone Marrow Transplant. 2006 May;37(10):967-76. doi: 10.1038/sj.bmt.1705358.
Adult mesenchymal stem cells (MSC) that are able to differentiate into various mesenchymal cell types are typically isolated from bone marrow, but their significant presence in human peripheral blood (PB) is controversial. Fibrin microbeads (FMB) that bind matrix-dependent cells were used to isolate MSC from the mononuclear fraction of mobilized PB of adult healthy human donors treated with a granulocyte colony-stimulating factor. Isolation by plastic adherence resulted in a negligible number of MSC in all samples tested, whereas FMB-based isolation yielded spindle-shaped cell samples that could further expand on plastic or on FMB in eight out of the 11 samples. The yield of these cells at days 17-18 after the harvest was approximately 0.5% of the initial cell number. The isolated cells were grown on plastic and characterized by FACS analysis and immunohistochemistry for specific markers. Following culturing and first passage, the FMB-isolated cells stained positive for mesenchymal stromal cell markers CD90 and CD105, expressed vimentin and fibronectin and were negative for hematopoietic markers CD45 and CD34. These cells could differentiate into osteoblasts, adipocytes and chondrocytes. This study indicates that FMB may have special advantage in isolating MSC from sources such as mobilized PB, where the number of such cells is scarce.
能够分化为多种间充质细胞类型的成人间充质干细胞（MSC）通常从骨髓中分离得到，但它们在人外周血（PB）中的大量存在存在争议。结合基质依赖性细胞的纤维蛋白微珠（FMB）被用于从接受粒细胞集落刺激因子治疗的成年健康人类供体的动员外周血单核细胞部分中分离MSC。通过塑料贴壁法分离，在所有测试样本中获得的MSC数量可忽略不计，而基于FMB的分离产生了纺锤形细胞样本，在11个样本中的8个样本中，这些细胞可以在塑料或FMB上进一步扩增。收获后第17 - 18天这些细胞的产量约为初始细胞数的0.5%。分离出的细胞在塑料上生长，并通过FACS分析和免疫组织化学检测特定标志物进行表征。培养和首次传代后，FMB分离的细胞间充质基质细胞标志物CD90和CD105染色呈阳性，表达波形蛋白和纤连蛋白，造血标志物CD45和CD34呈阴性。这些细胞可以分化为成骨细胞、脂肪细胞和软骨细胞。这项研究表明，FMB在从诸如动员外周血等此类细胞数量稀少的来源中分离MSC方面可能具有特殊优势。
