Characterization of mesenchymal stem cells isolated from murine bone marrow by negative selection
Characterization of mesenchymal stem cells isolated from murine bone marrow by negative selection
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DOI：
10.1002/jcb.10594
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发表时间：
2003-08-15
影响因子：
4
通讯作者：
Phinney, DG
中科院分区：
文献类型：
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作者：
Baddoo, M;Hill, K;Phinney, DG
间充质干细胞（MSCs）通常通过分离塑料贴壁的成纤维细胞样细胞组分从骨髓中富集。然而，从小鼠骨髓培养出的塑料贴壁细胞是成纤维细胞样细胞和造血细胞类型的混合物。在此我们报道一种基于免疫耗竭的可靠方法，用于从塑料贴壁的小鼠骨髓培养物中的造血细胞中分离成纤维细胞样细胞。免疫耗竭后的细胞表达抗原Sca - 1、CD29、CD44、CD81、CD106和干细胞标志物核干细胞因子（NST），但不表达CD11b、CD31、CD34、CD45、CD48、CD90、CD117、CD135或转录因子Oct - 4。它们还能够在体外分化为脂肪细胞、软骨细胞和成骨细胞，以及在体内分化为成骨细胞/骨细胞。因此，免疫耗竭产生了一个没有造血细胞和内皮细胞的细胞群体，其在表型和功能上等同于间充质干细胞。免疫耗竭后的细胞在培养中的群体倍增时间约为5 - 7天。生长不良是由于免疫耗竭导致许多参与细胞增殖和细胞周期进程的基因显著下调。免疫耗竭后的细胞暴露于成纤维细胞生长因子2（FGF2），而不是胰岛素样生长因子（IGF）、小鼠干细胞因子或白血病抑制因子（LIF），可显著提高其生长速率。此外，在FGF2存在的情况下，82%因免疫耗竭而下调的转录本保持不变。暴露于FGF2还可逆地抑制免疫耗竭后的细胞在体外分化为脂肪细胞、软骨细胞和成骨细胞的能力。因此，FGF2似乎作为一种有丝分裂原和自我维持因子，对通过负选择从骨髓中富集的小鼠间充质干细胞起作用。（C）2003威利 - 利斯公司
Mesenchymal stem cells (MSCs) are typically enriched from bone marrow via isolation of the plastic adherent, fibroblastoid cell fraction. However, plastic adherent cultures elaborated from murine bone marrow are an admixture of fibroblastoid and hematopoietic cell types. Here we report a reliable method based on immunodepletion to fractionate fibroblastoid cells from hematopoietic cells within plastic adherent murine marrow cultures. The immunodepleted cells expressed the antigens Sca-1, CD29, CD44, CD81, CD106, and the stem cell marker nucleostemin (NST) but not CD11b, CD31, CD34, CD45, CD48, CD90, CD117, CD135, or the transcription factor Oct-4. They were also capable of differentiating into adipocytes, chondrocytes, and osteoblasts in vitro as well as osteoblasts/osteocytes in vivo. Therefore, immunodepletion yields a cell population devoid of hematopoietic and endothelial cells that is phenotypically and functionally equivalent to MSCs. The immunodepleted cells exhibited a population doubling time of approximately 5-7 days in culture. Poor growth was due to the dramatic down regulation of many genes involved in cell proliferation and cell cycle progression as a result of immunodepletion. Exposure of immunodepleted cells to fibroblast growth factor 2 (FGF2) but not insulin-like growth factor (IGF), murine stem cell factor, or leukemia inhibitory factor (LIF) significantly increased their growth rate. Moreover, 82% of the transcripts down regulated by immunodepletion remain unaltered in the presence of FGF2. Exposure to the later also reversibly inhibited the ability of the immunodepleted cells to differentiate into adipocytes, chondrocytes, and osteoblasts in vitro. Therefore, FGF2 appears to function as a mitogen and self-maintenance factor for murine MSCs enriched from bone marrow by negative selection. (C) 2003 Wiley-Liss, Inc.