Monoclonal antibodies reactive with human osteogenic cell surface antigens
Monoclonal antibodies reactive with human osteogenic cell surface antigens
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DOI：
10.1016/s8756-3282(97)00127-0
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发表时间：
1997-09-01
期刊：
影响因子：
4.1
通讯作者：
Haynesworth, SE
中科院分区：
文献类型：
--
作者：
Bruder, SP;Horowitz, MC;Haynesworth, SE
针对成骨细胞表面的单克隆抗体（McAb）已被用于表征成骨谱系。鉴于针对正常人成骨细胞表面的探针的缺乏，我们试图产生可用于体内和体外研究的单克隆抗体。本研究以体外定向分化的人骨髓间充质干细胞（MSCs）免疫小鼠，制备了一系列抗早期成骨细胞表面抗原的单克隆抗体。经过对成骨细胞在体外分化的各个阶段的表面筛选，以及评估与人胎儿肢体的原位反应性，我们分离出三个杂交瘤细胞系，称为SB-10，SB-20和SB-21。成骨分化过程中的免疫细胞化学分析表明，SB-10与MSC和骨祖细胞反应，但一旦碱性磷酸酶（APase）表达，SB-10不再与细胞反应。流式细胞术证明SB-10在所有纯化的培养扩增的人MSC的表面上表达，从而提供了这些细胞是同质群体的进一步证据。相比之下，SB-20和SB-21不与原位祖细胞反应，但与AP酶阳性成骨细胞的子集结合。这些抗体都不能染色发育中骨切片中的终末分化骨细胞。此外，这些单克隆抗体在鸡、大鼠、兔、犬或牛骨样品中未观察到反应，尽管选择的皮肤外人组织进行了免疫染色。在检查的所有细胞和组织标本中，SB-20免疫染色与SB-21观察到的相同。我们已经使用这些单克隆抗体来完善我们对构成成骨细胞谱系的离散细胞转变的理解。我们提出了一个完善的模型，用于了解成骨细胞分化的基础上的命题，即顺序收购和特定的细胞表面分子的损失，可以用来定义成骨细胞谱系内的单个细胞的位置。(C)Elsevier Science Inc.出版。All rights reserved.
Monoclonal antibodies (McAbs) against the surface of osteoblastic cells have been used to characterize the osteogenic lineage. In view of the paucity of probes against the surface of normal human osteogenic cells, we sought to generate McAbs which could be used for both in vivo and in vitro studies. We raised a series of McAbs against early osteoblastic cell surface antigens by immunizing mice with human mesenchymal stem cells (MSCs) that had been directed into the osteogenic lineage in vitro. After screening against the surface of osteogenic cells at various stages of differentiation in vitro, as well as evaluating in situ reactivity with human fetal limbs, we isolated three hybridoma cell lines referred to as SB-10, SB-20, and SB-21. Immunocytochemical analyses during osteogenic differentiation demonstrate that SB-10 reacts with MSCs and osteoprogenitors, but no longer reacts with cells once alkaline phosphatase (APase) is expressed. Flow cytometry documents that SB-10 is expressed on the surface of all purified, culture-expanded human MSCs, thus providing further evidence that these cells are a homogeneous population. By contrast, SB-20 and SB-21 do not react with the progenitor cells in situ, but bind to a subset of the APase-positive osteoblasts. None of these antibodies stain terminally differentiated osteocytes in sections of developing bone. Furthermore, these McAbs were not observed to react in samples from chick, rat, rabbit, canine, or bovine bone, although selected extraskeletal human tissues were immunostained. In all cell and tissue specimens examined, SB-20 immunostaining is identical to that observed with SB-21. We have used these McAbs to refine our understanding of the discrete cellular transitions that constitute the osteogenic cell lineage. We suggest a refined model for understanding osteoblast differentiation that is based on the proposition that the sequential acquisition and loss of specific cell surface molecules can be used to define positions of individual cells within the osteogenic cell lineage. (C) 1997 by Elsevier Science Inc. All rights reserved.