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普通狨猴的造血祖细胞作为逆转录病毒和腺病毒载体基因转导的靶标。

Haematopoietic progenitor cells from the common marmoset as targets of gene transduction by retroviral and adenoviral vectors.

作者信息

Hibino H, Tani K, Sugiyama H, Suzuki S, Wu M S, Izawa K, Hase H, Nakazaki Y, Tanabe T, Ooi J, Izeki T, Tojo A, Saitoh I, Tanioka Y, Asano S

机构信息

Department of Hematology/Oncology, Institute of Medical Science, The University of Tokyo, Central Institute for Experimental Animals Laboratories Inc., Kawasaki, Kanagawa, Japan.

出版信息

Eur J Haematol. 2001 Apr;66(4):272-80. doi: 10.1034/j.1600-0609.2001.066004272.x.

Abstract

To establish a new non-human primate model for human cytokine and gene therapy, we characterized lymphocytes and haematopoietic progenitor cells of the small New World monkey, the common marmoset. We first assessed the reactions of marmoset bone marrow (BM) and peripheral blood (PB) cells to mouse anti-human monoclonal antibodies (mAbs) for the purpose of isolating marmoset lymphocytes and haematopoietic progenitor cells. Both cell fractions stained with CD4 and CD8 mAbs were identified as lymphocytes by cell proliferation assay and morphological examination. Myeloid-specific mAbs such as CD14 and CD33 did not react with marmoset BM and PB cells. No available CD34 and c-kit mAbs could be used to purify the marmoset haematopoietic progenitor cells. Furthermore, we studied the in vitro transduction of the bacterial beta-galactosidase (LacZ) gene into CFU-GM derived from marmoset BM using retroviral and adenoviral vectors. The transduction efficiency was increased by using a mixed culture system consisting of marmoset BM stromal cells and retroviral producer cells. It was also possible to transduce LacZ gene into marmoset haematopoietic progenitor cells with adenoviral vectors as well as retroviral vectors. The percentage of adenovirally transduced LacZ-positive clusters was 15% at day 4 (multiplicity of infection=200), but only 1-2% at day 14. The differential use of viral vector systems is to be recommended in targeting different diseases. Our results suggested that marmoset BM progenitor cells were available to examine the transduction efficiency of various viral vectors in vitro.

摘要

为建立一种用于人类细胞因子和基因治疗的新型非人类灵长类动物模型,我们对小型新大陆猴——普通狨猴的淋巴细胞和造血祖细胞进行了表征。我们首先评估了狨猴骨髓(BM)和外周血(PB)细胞对小鼠抗人单克隆抗体(mAb)的反应,目的是分离狨猴淋巴细胞和造血祖细胞。通过细胞增殖试验和形态学检查,用CD4和CD8单克隆抗体染色的两个细胞组分均被鉴定为淋巴细胞。髓系特异性单克隆抗体如CD14和CD33不与狨猴BM和PB细胞反应。没有可用的CD34和c-kit单克隆抗体可用于纯化狨猴造血祖细胞。此外,我们研究了使用逆转录病毒和腺病毒载体将细菌β-半乳糖苷酶(LacZ)基因体外转导至源自狨猴BM的CFU-GM中。通过使用由狨猴BM基质细胞和逆转录病毒生产细胞组成的混合培养系统,转导效率得以提高。使用腺病毒载体以及逆转录病毒载体也能够将LacZ基因转导至狨猴造血祖细胞中。腺病毒转导的LacZ阳性簇的百分比在第4天(感染复数=200)为15%,但在第14天仅为1-2%。在针对不同疾病时,建议差异使用病毒载体系统。我们的结果表明,狨猴BM祖细胞可用于体外检测各种病毒载体的转导效率。

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