Warrington A E, Barbarese E, Pfeiffer S E
Department of Microbiology, University of Connecticut School of Medicine, Farmington.
J Neurosci Res. 1993 Jan;34(1):1-13. doi: 10.1002/jnr.490340102.
The capacity of oligodendrocytes (OLs) and their progenitors to migrate, proliferate, and differentiate in vivo was evaluated by transplanting highly enriched populations of sequential stages of the OL lineage (A2B5+O4-, O4+GalC-, and GalC+) into the telencephalon of the hypomyelinating mouse, shiverer. The shiverer mouse neither expresses the major myelin basic protein (MBP) nor makes normal myelin due to a large deletion in the gene for MBP. Thirty days after transplantation, serial 225 micron sections of the host brain were immunostained with antiserum to MBP and analyzed by confocal microscopy. The presence of MBP+ patches of myelin in the otherwise MBP- host brain allowed a retrospective analysis of the myelinogenic activity of the transplanted progenitors cells. Both the extent of MBP+ myelin and the location of MBP+ structures relative to the initial site of cell deposition were highly dependent on the developmental stage of the transplanted cells. Specifically, A2B5+O4- OL progenitors migrated distances of > or = 600 microns and produced MBP+ patches in nearly every slice of the host brain. An average of over 250 separate patches were found per host brain, some of which had cross-sectional areas of > 250,000 microns2 containing as many as 60 MBP+ OL cell bodies, and with densities of myelination rivaling that of normal brain. In marked contrast, transplantation of O4+GalC- cells produced only small (1,000-25,000 microns2), scattered (25-40 per brain) patches of MBP+ myelin containing one to five cell bodies, all of which were within 50 microns of the needle track or the nearest ventricular surface. GalC+ cells produced MBP+ myelin at a level similar to that of O4+CalC- cells. These data suggest that the developmental transition of OL progenitors from the O4- to the O4+ phenotype is accompanied by a dramatic reduction in the innate capacity of the cells to migrate and survive in vivo. The use of developmentally identified, enriched populations of OL progenitor cells offers the opportunity for more precise analyses of transplantation and remyelination behavior, and relates to clinically relevant studies indicating that contaminant cell types can seriously interfere with the stable integration of donor tissue into the host.
通过将少突胶质细胞(OLs)及其祖细胞在不同发育阶段高度富集的细胞群体(A2B5 + O4 -、O4 + GalC -和GalC +)移植到髓鞘形成不足的颤抖小鼠的端脑中,评估其在体内迁移、增殖和分化的能力。颤抖小鼠由于髓鞘碱性蛋白(MBP)基因的大片段缺失,既不表达主要的髓鞘碱性蛋白,也不能形成正常的髓鞘。移植后30天,将宿主脑切成225微米厚的连续切片,用抗MBP血清进行免疫染色,并通过共聚焦显微镜分析。在原本缺乏MBP的宿主脑中出现MBP +髓鞘斑块,这使得对移植的祖细胞的髓鞘形成活性进行回顾性分析成为可能。MBP +髓鞘的范围以及MBP +结构相对于细胞沉积初始部位的位置,都高度依赖于移植细胞的发育阶段。具体而言,A2B5 + O4 -少突胶质祖细胞迁移的距离≥600微米,并且在宿主脑的几乎每一片切片中都产生了MBP +斑块。每个宿主脑中平均发现超过250个独立的斑块,其中一些斑块的横截面积>250,000平方微米,包含多达60个MBP +少突胶质细胞体,其髓鞘形成密度可与正常脑相媲美。与之形成鲜明对比的是,移植O4 + GalC -细胞仅产生小的(1,000 - 25,000平方微米)、分散的(每个脑25 - 40个)MBP +髓鞘斑块,每个斑块包含一到五个细胞体,所有这些细胞体都在针道或最近的脑室表面50微米范围内。GalC +细胞产生MBP +髓鞘的水平与O4 + CalC -细胞相似。这些数据表明,少突胶质祖细胞从O4 -表型向O4 +表型的发育转变伴随着细胞在体内迁移和存活的固有能力的显著降低。使用发育阶段明确、富集的少突胶质祖细胞群体为更精确地分析移植和髓鞘再生行为提供了机会,并且与临床相关研究相关,这些研究表明污染细胞类型会严重干扰供体组织在宿主体内的稳定整合。