Telomerized human bone marrow-derived cell clones maintain the phenotype of hematopoietic-supporting osteoblastic and myofibroblastic stromal cells after long-term culture
Telomerized human bone marrow-derived cell clones maintain the phenotype of hematopoietic-supporting osteoblastic and myofibroblastic stromal cells after long-term culture
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DOI：
10.1016/j.exphem.2005.09.004
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发表时间：
2005-12-01
影响因子：
2.6
通讯作者：
Niitsu, Y
中科院分区：
文献类型：
--
作者：
Kobune, M;Kato, J;Niitsu, Y
Objective.将端粒酶催化亚单位（TERT）基因转移到原代人基质细胞中可以延长其寿命。然而，原代人基质细胞实际上是由脂肪细胞，成肌脂细胞，成骨细胞等，我们的目的是调查的表型和造血支持的人端粒基质细胞（HTS）在克隆水平。材料和方法。我们通过有限稀释从HTS的亲本群体建立了HTS克隆（HTS-1至HTS-9）。通过与CD 34（+）细胞共培养检测HTS克隆的造血支持活性。HTS-1至HTS-3含有碱性磷酸酶（ALP）（+）细胞，HTS-4至HTS-9由ALP（+）和α-平滑肌肌动蛋白阳性细胞组成。尽管所有HTS克隆都表现出正常的生长动力学，但其中一个HTS克隆表现出染色体异常。此外，HTS细胞的亲本群体在培养4年后获得了增加的生长速率和锚定独立性。在所有HTS克隆中检测到造血生长因子，如干细胞因子、血管生成素-1和刺猬mRNA的表达。不同HTS克隆对造血祖细胞的支持程度不同，其中HTS-8的CD 34（+）细胞扩增水平最高。人端粒基质细胞克隆在长期培养后表现出造血支持成骨细胞和成肌纤维细胞的表型。HTS细胞的临床应用应受到限制，因为它们在hTERT基因转移后具有肿瘤转化的潜力。HTS细胞可用于分析人基质细胞造血支持的分子机制。(c)2005年国际实验血液学学会。爱思唯尔公司出版
Objective. Gene transfer of the telomerase catalytic subunit (TERT) into primary human stromal cells prolonged their lifespan. However, primary human stromal cells are actually composed of adipocytes, myolibroblasts, osteoblasts, etc. Our objective was to investigate the phenotype and hematopoietic-support of the human telomerized stromal cell (HTS) in clonal level.Materials and Methods. We established HTS clones (HTS-1 to HTS-9) from a parental population of HTSs by limiting dilution. Hematopoietic-supporting activity of the HTS clones was examined by coculturing with CD34(+) cells.Results. HTS-1 to HTS-3 contained alkaline phosphatase (ALP)(+) cells, and HTS-4 to HTS-9 were composed of both ALP(+) and alpha-smooth muscle actin-positive cells. Although all HTS clones exhibited normal growth kinetics, one of the HTS clones exhibited a chromosomal abnormality. Moreover, the parental population of the HTS cells acquired an increased growth rate and anchorage independence after 4 years of culturing. Expression of hematopoietic growth factors, such as stem cell factor, angiopoietin-1, and hedgehog mRNA was detected in all HTS clones. The degree of hematopoietic progenitor support differed between the HTS clones, and the expansion level of CD34(+) cells was the highest in HTS-8.Conclusion. Human telomerized stromal cell clones exhibited the phenotype of hematopoietic supporting osteoblastic and myofibroblastic cells after long-term culture. Clinical application of HTS cells should be limited because of their potential for neoplastic transformation after hTERT gene transfer. HTS cells may be useful for analyzing the molecular mechanism of hematopoietic support of human stromal cells. (c) 2005 International Society for Experimental Hematology. Published by Elsevier Inc.