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通过抗c-kit(CD117)单克隆抗体2B8/BM鉴定的猪骨髓祖细胞的特性

Characterisation of porcine bone marrow progenitor cells identified by the anti-c-kit (CD117) monoclonal antibody 2B8/BM.

作者信息

Pérez C, Moreno S, Summerfield A, Domenech N, Alvarez B, Correa C, Alonso F, Ezquerra A, Domínguez J, Revilla C

机构信息

Dpto. Biotecnología, Instituto Nacional de Investigación y Tecnología Agraria y Alimentaria (INIA), 28040, Madrid, Spain.

出版信息

J Immunol Methods. 2007 Apr 10;321(1-2):70-9. doi: 10.1016/j.jim.2007.01.003. Epub 2007 Feb 5.

Abstract

c-kit (CD117) plays an important role in the early stages of haematopoiesis. Previous studies of porcine haematopoietic stem cells have relied for their identification on the use of the c-kit ligand stem cell factor. Here, we describe a new mAb, 2B8/BM, that recognizes a 155-kDa protein expressed on a small subset (2-8%) of bone marrow haematopoietic cells. 2B8/BM(+) cells have a blast appearance, and are mostly negative for lineage-specific markers or express low levels of CD172a or SLA-II. In in vitro colony-forming unit assays these cells were able to give rise to erythroid and myeloid colonies. Altogether these data suggested that the 2B8/BM antigen might be the porcine orthologue of the human c-kit. This specificity was confirmed by the binding of mAb 2B8/BM to CHO cells transfected with a plasmid encoding the porcine c-kit ectodomain. This antibody can facilitate the isolation and enrichment of porcine stem cells to be used in procedures aimed to induce xenograft tolerance or to test their potential to repair damaged tissues and organs.

摘要

c-kit(CD117)在造血早期阶段发挥着重要作用。先前对猪造血干细胞的研究依赖于使用c-kit配体干细胞因子来进行鉴定。在此,我们描述了一种新的单克隆抗体2B8/BM,它可识别在一小部分(2%-8%)骨髓造血细胞上表达的一种155 kDa的蛋白质。2B8/BM(+)细胞呈原始细胞形态,大多对谱系特异性标志物呈阴性,或低水平表达CD172a或SLA-II。在体外集落形成单位试验中,这些细胞能够产生红系和髓系集落。总体而言,这些数据表明2B8/BM抗原可能是人类c-kit的猪同源物。单克隆抗体2B8/BM与用编码猪c-kit胞外结构域的质粒转染的CHO细胞结合,证实了这种特异性。这种抗体有助于分离和富集猪干细胞,用于旨在诱导异种移植耐受或测试其修复受损组织和器官潜力的程序中。

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